239,097 results on '"Ligands"'
Search Results
2. Proximity Graph Networks: Predicting Ligand Affinity with Message Passing Neural Networks.
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Gale-Day, Zachary, Shub, Laura, Chuang, Kangway, and Keiser, Michael
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Ligands ,Neural Networks ,Computer ,Proteins ,Molecular Docking Simulation ,Protein Binding - Abstract
Message passing neural networks (MPNNs) on molecular graphs generate continuous and differentiable encodings of small molecules with state-of-the-art performance on protein-ligand complex scoring tasks. Here, we describe the proximity graph network (PGN) package, an open-source toolkit that constructs ligand-receptor graphs based on atom proximity and allows users to rapidly apply and evaluate MPNN architectures for a broad range of tasks. We demonstrate the utility of PGN by introducing benchmarks for affinity and docking score prediction tasks. Graph networks generalize better than fingerprint-based models and perform strongly for the docking score prediction task. Overall, MPNNs with proximity graph data structures augment the prediction of ligand-receptor complex properties when ligand-receptor data are available.
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- 2024
3. Engineering programmable material-to-cell pathways via synthetic notch receptors to spatially control differentiation in multicellular constructs.
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Garibyan, Mher, Hoffman, Tyler, Makaske, Thijs, Do, Stephanie, Wu, Yifan, Williams, Brian, March, Alexander, Cho, Nathan, Pedroncelli, Nicolas, Lima, Ricardo, Soto, Jennifer, Jackson, Brooke, Santoso, Jeffrey, Khademhosseini, Ali, Thomson, Matt, Li, Song, McCain, Megan, and Morsut, Leonardo
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Receptors ,Notch ,Cell Differentiation ,Tissue Engineering ,Animals ,Humans ,Signal Transduction ,Mice ,Extracellular Matrix ,Fibroblasts ,Extracellular Matrix Proteins ,Ligands ,Tissue Scaffolds ,Muscle ,Skeletal ,Endothelial Cells ,HEK293 Cells - Abstract
Synthetic Notch (synNotch) receptors are genetically encoded, modular synthetic receptors that enable mammalian cells to detect environmental signals and respond by activating user-prescribed transcriptional programs. Although some materials have been modified to present synNotch ligands with coarse spatial control, applications in tissue engineering generally require extracellular matrix (ECM)-derived scaffolds and/or finer spatial positioning of multiple ligands. Thus, we develop here a suite of materials that activate synNotch receptors for generalizable engineering of material-to-cell signaling. We genetically and chemically fuse functional synNotch ligands to ECM proteins and ECM-derived materials. We also generate tissues with microscale precision over four distinct reporter phenotypes by culturing cells with two orthogonal synNotch programs on surfaces microcontact-printed with two synNotch ligands. Finally, we showcase applications in tissue engineering by co-transdifferentiating fibroblasts into skeletal muscle or endothelial cell precursors in user-defined micropatterns. These technologies provide avenues for spatially controlling cellular phenotypes in mammalian tissues.
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- 2024
4. Structure-based discovery of CFTR potentiators and inhibitors.
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Liu, Fangyu, Kaplan, Anat, Levring, Jesper, Einsiedel, Jürgen, Tiedt, Stephanie, Distler, Katharina, Omattage, Natalie, Kondratov, Ivan, Moroz, Yurii, Pietz, Harlan, Irwin, John, Gmeiner, Peter, Shoichet, Brian, and Chen, Jue
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ABC transporter ,anion channel ,inhibitors ,large-scale docking ,ligand discovery ,potentiators ,Cystic Fibrosis Transmembrane Conductance Regulator ,Humans ,Molecular Docking Simulation ,Cystic Fibrosis ,Aminophenols ,Drug Discovery ,Cryoelectron Microscopy ,Quinolones ,Allosteric Site ,Animals ,Ligands - Abstract
The cystic fibrosis transmembrane conductance regulator (CFTR) is a crucial ion channel whose loss of function leads to cystic fibrosis, whereas its hyperactivation leads to secretory diarrhea. Small molecules that improve CFTR folding (correctors) or function (potentiators) are clinically available. However, the only potentiator, ivacaftor, has suboptimal pharmacokinetics and inhibitors have yet to be clinically developed. Here, we combine molecular docking, electrophysiology, cryo-EM, and medicinal chemistry to identify CFTR modulators. We docked ∼155 million molecules into the potentiator site on CFTR, synthesized 53 test ligands, and used structure-based optimization to identify candidate modulators. This approach uncovered mid-nanomolar potentiators, as well as inhibitors, that bind to the same allosteric site. These molecules represent potential leads for the development of more effective drugs for cystic fibrosis and secretory diarrhea, demonstrating the feasibility of large-scale docking for ion channel drug discovery.
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- 2024
5. Reliable ligand discrimination in stochastic multistep kinetic proofreading: First passage time vs. product counting strategies.
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Li, Xiangting and Chou, Tom
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Kinetics ,Stochastic Processes ,Ligands ,Receptors ,Antigen ,T-Cell ,Signal Transduction ,Computational Biology ,Models ,Biological ,Humans ,DNA Replication - Abstract
Cellular signaling, crucial for biological processes like immune response and homeostasis, relies on specificity and fidelity in signal transduction to accurately respond to stimuli amidst biological noise. Kinetic proofreading (KPR) is a key mechanism enhancing signaling specificity through time-delayed steps, although its effectiveness is debated due to intrinsic noise potentially reducing signal fidelity. In this study, we reformulate the theory of kinetic proofreading (KPR) by convolving multiple intermediate states into a single state and then define an overall processing time required to traverse these states. This simplification allows us to succinctly describe kinetic proofreading in terms of a single waiting time parameter, facilitating a more direct evaluation and comparison of KPR performance across different biological contexts such as DNA replication and T cell receptor (TCR) signaling. We find that loss of fidelity for longer proofreading steps relies on the specific strategy of information extraction and show that in the first-passage time (FPT) discrimination strategy, longer proofreading steps can exponentially improve the accuracy of KPR at the cost of speed. Thus, KPR can still be an effective discrimination mechanism in the high noise regime. However, in a product concentration-based discrimination strategy, longer proofreading steps do not necessarily lead to an increase in performance. However, by introducing activation thresholds on product concentrations, can we decompose the product-based strategy into a series of FPT-based strategies to better resolve the subtleties of KPR-mediated product discrimination. Our findings underscore the importance of understanding KPR in the context of how information is extracted and processed in the cell.
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- 2024
6. The landscape of cancer-rewired GPCR signaling axes.
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Arora, Chakit, Matic, Marin, Bisceglia, Luisa, Di Chiaro, Pierluigi, De Oliveira Rosa, Natalia, Carli, Francesco, Clubb, Lauren, Nemati Fard, Lorenzo, Kargas, Giorgos, Diaferia, Giuseppe, Vukotic, Ranka, Licata, Luana, Wu, Guanming, Natoli, Gioacchino, Gutkind, J, and Raimondi, Francesco
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GPCR ,cancer ,cancer cell lines ,cell-cell communication ,drug repurposing ,personalized medicine ,signaling network ,survival analysis ,transcriptomics ,Humans ,Receptors ,G-Protein-Coupled ,Neoplasms ,Signal Transduction ,Ligands ,Gene Expression Regulation ,Neoplastic - Abstract
We explored the dysregulation of G-protein-coupled receptor (GPCR) ligand systems in cancer transcriptomics datasets to uncover new therapeutics opportunities in oncology. We derived an interaction network of receptors with ligands and their biosynthetic enzymes. Multiple GPCRs are differentially regulated together with their upstream partners across cancer subtypes and are associated to specific transcriptional programs and to patient survival patterns. The expression of both receptor-ligand (or enzymes) partners improved patient stratification, suggesting a synergistic role for the activation of GPCR networks in modulating cancer phenotypes. Remarkably, we identified many such axes across several cancer molecular subtypes, including many involving receptor-biosynthetic enzymes for neurotransmitters. We found that GPCRs from these actionable axes, including, e.g., muscarinic, adenosine, 5-hydroxytryptamine, and chemokine receptors, are the targets of multiple drugs displaying anti-growth effects in large-scale, cancer cell drug screens, which we further validated. We have made the results generated in this study freely available through a webapp (gpcrcanceraxes.bioinfolab.sns.it).
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- 2024
7. X-ray crystal structure of a designed rigidified imaging scaffold in the ligand-free conformation.
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Agdanowski, Matthew, Castells-Graells, Roger, Sawaya, Michael, Cascio, Duilio, Yeates, Todd, and Arbing, Mark
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DARPins ,imaging scaffolds ,protein cages ,protein design ,Crystallography ,X-Ray ,Models ,Molecular ,Ankyrin Repeat ,Cryoelectron Microscopy ,Ligands ,Protein Conformation ,Protein Binding ,Gene Expression - Abstract
Imaging scaffolds composed of designed protein cages fused to designed ankyrin repeat proteins (DARPins) have enabled the structure determination of small proteins by cryogenic electron microscopy (cryo-EM). One particularly well characterized scaffold type is a symmetric tetrahedral assembly composed of 24 subunits, 12 A and 12 B, which has three cargo-binding DARPins positioned on each vertex. Here, the X-ray crystal structure of a representative tetrahedral scaffold in the apo state is reported at 3.8 Å resolution. The X-ray crystal structure complements recent cryo-EM findings on a closely related scaffold, while also suggesting potential utility for crystallographic investigations. As observed in this crystal structure, one of the three DARPins, which serve as modular adaptors for binding diverse `cargo proteins, present on each of the vertices is oriented towards a large solvent channel. The crystal lattice is unusually porous, suggesting that it may be possible to soak crystals of the scaffold with small (≤30 kDa) protein cargo ligands and subsequently determine cage-cargo structures via X-ray crystallography. The results suggest the possibility that cryo-EM scaffolds may be repurposed for structure determination by X-ray crystallography, thus extending the utility of electron-microscopy scaffold designs for alternative structural biology applications.
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- 2024
8. Real-World Experience with 177Lu-PSMA-617 Radioligand Therapy After Food and Drug Administration Approval
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Moradi Tuchayi, Abuzar, Yadav, Surekha, Jiang, Fei, Kim, Sarasa T, Saelee, Rachelle K, Morley, Amanda, Juarez, Roxanna, Lawhn-Heath, Courtney, Wang, Yingbing, de Kouchkovsky, Ivan, and Hope, Thomas A
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Biomedical and Clinical Sciences ,Clinical Sciences ,Urologic Diseases ,Good Health and Well Being ,Humans ,Male ,Lutetium ,Aged ,Heterocyclic Compounds ,1-Ring ,Retrospective Studies ,Dipeptides ,Middle Aged ,United States ,United States Food and Drug Administration ,Prostate-Specific Antigen ,Aged ,80 and over ,Drug Approval ,Ligands ,Treatment Outcome ,Prostatic Neoplasms ,Radiopharmaceuticals ,177Lu-PSMA-617 ,genitourinary ,oncology ,radionuclide therapy ,toxicities ,Nuclear Medicine & Medical Imaging ,Clinical sciences - Abstract
We report our initial real-world experience with 177Lu-PSMA-617 radioligand therapy. Methods: We performed a retrospective review of patients treated with 177Lu-PSMA-617. Pretreatment PSMA PET, laboratory findings, overall survival, a fall in prostate-specific antigen by 50% (PSA50), and toxicities were evaluated. Results: Ninety-nine patients were included. Sixty patients achieved a PSA50. Seven of 18 (39%) patients who did not meet the TheraP PSMA imaging criteria achieved a PSA50. Nineteen of 31 (61%) patients who did not meet the VISION laboratory criteria achieved a PSA50. Sixty-three patients had a delay or stoppage in therapy, which was due to a good response in 19 patients and progressive disease in 14 patients. Of 10 patients with a good response who restarted treatment, 9 subsequently achieved a PSA50 on retreatment. The most common toxicities were anemia (33%) and thrombocytopenia (21%). Conclusion: At our center, patients who did not meet the TheraP PSMA imaging criteria or the VISION laboratory criteria benefited from 177Lu-PSMA-617 radioligand therapy.
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- 2024
9. Axon‐derived PACSIN1 binds to the Schwann cell survival receptor, LRP1, and transactivates TrkC to promote gliatrophic activities
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Martellucci, Stefano, Flütsch, Andreas, Carter, Mark, Norimoto, Masaki, Pizzo, Donald, Mantuano, Elisabetta, Sadri, Mahrou, Wang, Zixuan, Chillin‐Fuentes, Daisy, Rosenthal, Sara Brin, Azmoon, Pardis, Gonias, Steven L, and Campana, Wendy M
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Biomedical and Clinical Sciences ,Neurosciences ,Regenerative Medicine ,Physical Injury - Accidents and Adverse Effects ,Aetiology ,2.1 Biological and endogenous factors ,Neurological ,Animals ,Mice ,Rats ,Axons ,Cell Survival ,Cells ,Cultured ,Ligands ,Rats ,Sprague-Dawley ,Receptor Protein-Tyrosine Kinases ,Schwann Cells ,Humans ,Adaptor Proteins ,Signal Transducing ,Recombinant Proteins ,axon glia interaction ,cell signaling ,injury-repair ,LRP1 ,Schwann cells ,Neurology & Neurosurgery - Abstract
Schwann cells (SCs) undergo phenotypic transformation and then orchestrate nerve repair following PNS injury. The ligands and receptors that activate and sustain SC transformation remain incompletely understood. Proteins released by injured axons represent important candidates for activating the SC Repair Program. The low-density lipoprotein receptor-related protein-1 (LRP1) is acutely up-regulated in SCs in response to injury, activating c-Jun, and promoting SC survival. To identify novel LRP1 ligands released in PNS injury, we applied a discovery-based approach in which extracellular proteins in the injured nerve were captured using Fc-fusion proteins containing the ligand-binding motifs of LRP1 (CCR2 and CCR4). An intracellular neuron-specific protein, Protein Kinase C and Casein Kinase Substrate in Neurons (PACSIN1) was identified and validated as an LRP1 ligand. Recombinant PACSIN1 activated c-Jun and ERK1/2 in cultured SCs. Silencing Lrp1 or inhibiting the LRP1 cell-signaling co-receptor, the NMDA-R, blocked the effects of PACSIN1 on c-Jun and ERK1/2 phosphorylation. Intraneural injection of PACSIN1 into crush-injured sciatic nerves activated c-Jun in wild-type mice, but not in mice in which Lrp1 is conditionally deleted in SCs. Transcriptome profiling of SCs revealed that PACSIN1 mediates gene expression events consistent with transformation to the repair phenotype. PACSIN1 promoted SC migration and viability following the TNFα challenge. When Src family kinases were pharmacologically inhibited or the receptor tyrosine kinase, TrkC, was genetically silenced or pharmacologically inhibited, PACSIN1 failed to induce cell signaling and prevent SC death. Collectively, these studies demonstrate that PACSIN1 is a novel axon-derived LRP1 ligand that activates SC repair signaling by transactivating TrkC.
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- 2024
10. Model of P-Glycoprotein Ligand Binding and Validation with Efflux Substrate Matched Pairs.
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Conrad, Jay, Vaz, Roy, and Paras, Nick
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Humans ,ATP Binding Cassette Transporter ,Subfamily B ,Member 1 ,Ligands ,ATP Binding Cassette Transporter ,Subfamily B ,Blood-Brain Barrier ,Neoplasms - Abstract
The blood-brain barrier (BBB) poses a significant obstacle in developing therapeutics for neurodegenerative diseases and central nervous system (CNS) disorders. P-glycoprotein (P-gp), a multidrug resistance protein, is a critical gatekeeper in the BBB and plays a role in cancer chemoresistance. This paper uses cryo-EM P-gp structures as starting points with an induced fit docking (IFD) model to evaluate 19 pairs of compounds with known P-gp efflux data. The study reveals significant differences in binding energy and sheds light on structural modifications impact on efflux properties. In the cases examined, fluorine incorporation influences the efflux by altering the molecular conformation rather than proximal heteroatom basicity. Although there are limitations in addressing covalent interactions or when binding extends into the more flexible vestibule region of the protein, the results provide valuable insights and potential strategies to overcome P-gp efflux, contributing to the advancement of drug development for both CNS disorders and cancer therapies.
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- 2024
11. Co-imaging of RelA and c-Rel reveals features of NF-κB signaling for ligand discrimination.
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Rahman, Shah, Singh, Apeksha, Lowe, Sarina, Aqdas, Mohammad, Jiang, Kevin, Vaidehi Narayanan, Haripriya, Hoffmann, Alexander, and Sung, Myong-Hee
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CP: Molecular biology ,NF-κB ,RelA ,c-Rel ,endogenous knockin ,fluorescent fusion reporter mice ,inflammatory signaling ,live microscopy ,macrophages ,mathematical modeling ,Mice ,Animals ,NF-kappa B ,Ligands ,Proto-Oncogene Proteins c-rel ,Transcription Factor RelA ,Signal Transduction ,Macrophages - Abstract
Individual cell sensing of external cues has evolved through the temporal patterns in signaling. Since nuclear factor κB (NF-κB) signaling dynamics have been examined using a single subunit, RelA, it remains unclear whether more information might be transmitted via other subunits. Using NF-κB double-knockin reporter mice, we monitored both canonical NF-κB subunits, RelA and c-Rel, simultaneously in single macrophages by quantitative live-cell imaging. We show that signaling features of RelA and c-Rel convey more information about the stimuli than those of either subunit alone. Machine learning is used to predict the ligand identity accurately based on RelA and c-Rel signaling features without considering the co-activated factors. Ligand discrimination is achieved through selective non-redundancy of RelA and c-Rel signaling dynamics, as well as their temporal coordination. These results suggest a potential role of c-Rel in fine-tuning immune responses and highlight the need for approaches that will elucidate the mechanisms regulating NF-κB subunit specificity.
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- 2024
12. Engineering a Programmed Death-Ligand 1-Targeting Monobody Via Directed Evolution for SynNotch-Gated Cell Therapy.
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Zhu, Linshan, Man, Chi-Wei, Harrison, Reed, Wu, Zhuohang, Limsakul, Praopim, Peng, Qin, Hashimoto, Matthew, Mamaril, Anthony, Xu, Hongquan, Liu, Longwei, and Wang, Yingxiao
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CAR T cell therapy ,Directed evolution ,Monobody ,PD-L1 ,SynNotch ,Yeast surface display ,Humans ,Mice ,Animals ,Receptors ,Antigen ,T-Cell ,B7-H1 Antigen ,Ligands ,Cell Line ,Tumor ,T-Lymphocytes ,Immunotherapy ,Adoptive - Abstract
Programmed death-ligand 1 (PD-L1) is a promising target for cancer immunotherapy due to its ability to inhibit T cell activation; however, its expression on various noncancer cells may cause on-target off-tumor toxicity when designing PD-L1-targeting Chimeric Antigen Receptor (CAR) T cell therapies. Combining rational design and directed evolution of the human fibronectin-derived monobody scaffold, PDbody was engineered to bind to PD-L1 with a preference for a slightly lower pH, which is typical in the tumor microenvironment. PDbody was further utilized as a CAR to target the PD-L1-expressing triple negative MDA-MB-231 breast cancer cell line. To mitigate on-target off-tumor toxicity associated with targeting PD-L1, a Cluster of Differentiation 19 (CD19)-recognizing SynNotch IF THEN gate was integrated into the system. This CD19-SynNotch PDbody-CAR system was then expressed in primary human T cells to target CD19-expressing MDA-MB-231 cancer cells. These CD19-SynNotch PDbody-CAR T cells demonstrated both specificity and efficacy in vitro, accurately eradicating cancer targets in cytotoxicity assays. Moreover, in an in vivo bilateral murine tumor model, they exhibited the capability to effectively restrain tumor growth. Overall, CD19-SynNotch PDbody-CAR T cells represent a distinct development over previously published designs due to their increased efficacy, proliferative capability, and mitigation of off-tumor toxicity for solid tumor treatment.
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- 2024
13. PopShift: A Thermodynamically Sound Approach to Estimate Binding Free Energies by Accounting for Ligand-Induced Population Shifts from a Ligand-Free Markov State Model.
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Smith, Louis, Novak, Borna, Osato, Meghan, Bowman, Gregory, and Mobley, David
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Protein Binding ,Ligands ,Proteins ,Entropy ,Protein Conformation ,Thermodynamics ,Binding Sites - Abstract
Obtaining accurate binding free energies from in silico screens has been a long-standing goal for the computational chemistry community. However, accuracy and computational cost are at odds with one another, limiting the utility of methods that perform this type of calculation. Many methods achieve massive scale by explicitly or implicitly assuming that the target protein adopts a single structure, or undergoes limited fluctuations around that structure, to minimize computational cost. Others simulate each protein-ligand complex of interest, accepting lower throughput in exchange for better predictions of binding affinities. Here, we present the PopShift framework for accounting for the ensemble of structures a protein adopts and their relative probabilities. Protein degrees of freedom are enumerated once, and then arbitrarily many molecules can be screened against this ensemble. Specifically, we use Markov state models (MSMs) as a compressed representation of a proteins thermodynamic ensemble. We start with a ligand-free MSM and then calculate how addition of a ligand shifts the populations of each protein conformational state based on the strength of the interaction between that protein conformation and the ligand. In this work we use docking to estimate the affinity between a given protein structure and ligand, but any estimator of binding affinities could be used in the PopShift framework. We test PopShift on the classic benchmark pocket T4 Lysozyme L99A. We find that PopShift is more accurate than common strategies, such as docking to a single structure and traditional ensemble docking─producing results that compare favorably with alchemical binding free energy calculations in terms of RMSE but not correlation─and may have a more favorable computational cost profile in some applications. In addition to predicting binding free energies and ligand poses, PopShift also provides insight into how the probability of different protein structures is shifted upon addition of various concentrations of ligand, providing a platform for predicting affinities and allosteric effects of ligand binding. Therefore, we expect PopShift will be valuable for hit finding and for providing insight into phenomena like allostery.
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- 2024
14. DockOpt: A Tool for Automatic Optimization of Docking Models.
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Knight, Ian, Mailhot, Olivier, Tang, Khanh, and Irwin, John
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Molecular Docking Simulation ,Prospective Studies ,Proteins ,Ligands ,Benchmarking ,Protein Binding - Abstract
Molecular docking is a widely used technique for leveraging protein structure for ligand discovery, but it remains difficult to utilize due to limitations that have not been adequately addressed. Despite some progress toward automation, docking still requires expert guidance, hindering its adoption by a broader range of investigators. To make docking more accessible, we developed a new utility called DockOpt, which automates the creation, evaluation, and optimization of docking models prior to their deployment in large-scale prospective screens. DockOpt outperforms our previous automated pipeline across all 43 targets in the DUDE-Z benchmark data set, and the generated models for 84% of targets demonstrate sufficient enrichment to warrant their use in prospective screens, with normalized LogAUC values of at least 15%. DockOpt is available as part of the Python package Pydock3 included in the UCSF DOCK 3.8 distribution, which is available for free to academic researchers at https://dock.compbio.ucsf.edu and free for everyone upon registration at https://tldr.docking.org.
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- 2024
15. Gold(I) ion and the phosphine ligand are necessary for the anti-Toxoplasma gondii activity of auranofin.
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Ma, C, Tirtorahardjo, J, Schweizer, S, Zhang, J, Fang, Z, Xing, L, Xu, M, Herman, D, Kleinman, M, McCullough, B, Barrios, A, and Andrade, Rosa
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Toxoplasma ,auranofin ,drug discovery ,gold ,repurposing ,Humans ,Auranofin ,Gold ,Toxoplasma ,Ligands ,Aurothioglucose ,Arthritis ,Rheumatoid ,Gold Sodium Thiomalate ,Toxoplasmosis ,Antiparasitic Agents ,Phosphines - Abstract
Auranofin, an FDA-approved drug for rheumatoid arthritis, has emerged as a promising antiparasitic medication in recent years. The gold(I) ion in auranofin is postulated to be responsible for its antiparasitic activity. Notably, aurothiomalate and aurothioglucose also contain gold(I), and, like auranofin, they were previously used to treat rheumatoid arthritis. Whether they have antiparasitic activity remains to be elucidated. Herein, we demonstrated that auranofin and similar derivatives, but not aurothiomalate and aurothioglucose, inhibited the growth of Toxoplasma gondii in vitro. We found that auranofin affected the T. gondii biological cycle (lytic cycle) by inhibiting T. gondiis invasion and triggering its egress from the host cell. However, auranofin could not prevent parasite replication once T. gondii resided within the host. Auranofin treatment induced apoptosis in T. gondii parasites, as demonstrated by its reduced size and elevated phosphatidylserine externalization (PS). Notably, the gold from auranofin enters the cytoplasm of T. gondii, as demonstrated by scanning transmission electron microscopy-energy dispersive X-ray spectroscopy (STEM-EDS) and Inductively Coupled Plasma-Mass Spectrometry (ICP-MS).IMPORTANCEToxoplasmosis, caused by Toxoplasma gondii, is a devastating disease affecting the brain and the eyes, frequently affecting immunocompromised individuals. Approximately 60 million people in the United States are already infected with T. gondii, representing a population at-risk of developing toxoplasmosis. Recent advances in treating cancer, autoimmune diseases, and organ transplants have contributed to this at-risk populations exponential growth. Paradoxically, treatments for toxoplasmosis have remained the same for more than 60 years, relying on medications well-known for their bone marrow toxicity and allergic reactions. Discovering new therapies is a priority, and repurposing FDA-approved drugs is an alternative approach to speed up drug discovery. Herein, we report the effect of auranofin, an FDA-approved drug, on the biological cycle of T. gondii and how both the phosphine ligand and the gold molecule determine the anti-parasitic activity of auranofin and other gold compounds. Our studies would contribute to the pipeline of candidate anti-T. gondii agents.
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- 2024
16. Phase 1/2 study of monalizumab plus durvalumab in patients with advanced solid tumors.
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Patel, Sandip, Alonso-Gordoa, Teresa, Banerjee, Susana, Wang, Ding, Naidoo, Jarushka, Standifer, Nathan, Palmer, Doug, Cheng, Lin-Yang, Kourtesis, Panagiotis, Ascierto, Maria, Das, Mayukh, Diamond, Jennifer, Hellmann, Matthew, and Carneiro, Benedito
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Adaptive Immunity ,Immunity ,Innate ,Natural Killer T-Cells ,Programmed Cell Death 1 Receptor ,Tumor Microenvironment ,Female ,Humans ,Adolescent ,Adult ,Carcinoma ,Non-Small-Cell Lung ,Ligands ,Lung Neoplasms ,Tumor Microenvironment ,Antibodies ,Monoclonal ,Antibodies ,Monoclonal ,Humanized - Abstract
BACKGROUND: The combination of monalizumab (anti-NKG2A/CD94) and durvalumab (anti-programmed death ligand-1) may promote antitumor immunity by targeting innate and adaptive immunity. This phase 1/2 study of monalizumab and durvalumab evaluated safety, antitumor activity, and pharmacodynamics in patients with advanced solid tumors. MAIN BODY: Immunotherapy-naïve patients aged ≥18 years with advanced disease, Eastern Cooperative Oncology Group performance status of 0-1, and 1-3 prior lines of systemic therapy in the recurrent/metastatic setting were enrolled. In part 1 (dose escalation), patients received durvalumab 1500 mg every 4 weeks (Q4W) with increasing doses of monalizumab Q2W/Q4W (n=15). Dose expansion in part 1 included patients with cervical cancer (n=15; durvalumab 1500 mg Q4W and monalizumab 750 mg Q2W) or metastatic microsatellite stable (MSS)-colorectal cancer (CRC) (n=15; durvalumab 1500 mg Q4W and monalizumab 750 mg Q4W). In part 2 (dose expansion), patients with MSS-CRC (n=40), non-small cell lung cancer (NSCLC; n=20), MSS-endometrial cancer (n=40), or ovarian cancer (n=40) received durvalumab 1500 mg Q4W and monalizumab 750 mg Q2W. The primary endpoint was safety. Secondary endpoints included antitumor activity per Response Evaluation Criteria In Solid Tumors version 1.1 (RECIST v1.1). Exploratory analyses included assessment of T-cell and natural killer (NK) cell activation and proliferation in peripheral blood and the tumor microenvironment (TME). The study enrolled 185 patients (part 1, 45; part 2, 140). No dose-limiting toxicities were observed and the maximum tolerated dose was not reached. In part 2, the most common treatment-related adverse events were fatigue (12.1%), asthenia (9.3%), diarrhea (9.3%), pruritus (7.9%), and pyrexia (7.1%). In the expansion cohorts, response rates were 0% (cervical), 7.7% (MSS-CRC), 10% (NSCLC), 5.4% (ovarian), and 0% (MSS-endometrial). Sustained NK cell activation, CD8+ T-cell proliferation, increased serum levels of CXCL10 (C-X-C motif chemokine ligand 10) and CXCL11, and increased tumor infiltration of CD8+ and granzyme B+ cells were observed. CONCLUSIONS: Although efficacy was modest, monalizumab plus durvalumab was well tolerated and encouraging immune activation was observed in the peripheral blood and TME. TRIAL REGISTRATION NUMBER: NCT02671435.
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- 2024
17. Carbon monoxide poisoning: A problem uniquely suited to a medicinal inorganic chemistry solution
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Parker, A Leila and Johnstone, Timothy C
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Inorganic Chemistry ,Chemical Sciences ,Humans ,Carbon Monoxide Poisoning ,Antidotes ,Ligands ,Carbon Monoxide ,Oxygen ,Chemistry ,Inorganic ,Antidote ,Bis-pocket porphyrin ,Carbon monoxide poisoning ,HemoCD ,Neuroglobin ,Porphyrin ,Theoretical and Computational Chemistry ,Other Chemical Sciences ,Inorganic & Nuclear Chemistry ,Inorganic chemistry - Abstract
Carbon monoxide poisoning is one of the most common forms of poisoning in the world. Although the primary mode of treatment, oxygen therapy, is highly effective in many cases, there are instances in which it is inadequate or inappropriate. Whereas oxygen therapy relies on high levels of a low-affinity ligand (O2) to displace a high-affinity ligand (CO) from metalloproteins, an antidote strategy relies on introducing a molecule with a higher affinity for CO than native proteins (Kantidote,CO > Kprotein,CO). Based on the fundamental chemistry of CO, such an antidote is most likely required to be an inorganic compound featuring an electron-rich transition metal. A review is provided of the protein-, supramolecular complex-, and small molecule-based CO poisoning antidote platforms that are currently under investigation.
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- 2024
18. Docking for Molecules That Bind in a Symmetric Stack with SymDOCK.
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Smith, Matthew, Knight, Ian, Kormos, Rian, Pepe, Joseph, Kunach, Peter, Diamond, Marc, Shahmoradian, Sarah, Irwin, John, DeGrado, William, and Shoichet, Brian
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Prospective Studies ,Ligands ,Retrospective Studies ,Proteins ,Molecular Docking Simulation ,Protein Binding ,Binding Sites - Abstract
Discovering ligands for amyloid fibrils, such as those formed by the tau protein, is an area of great current interest. In recent structures, ligands bind in stacks in the tau fibrils to reflect the rotational and translational symmetry of the fibril itself; in these structures, the ligands make few interactions with the protein but interact extensively with each other. To exploit this symmetry and stacking, we developed SymDOCK, a method to dock molecules that follow the proteins symmetry. For each prospective ligand pose, we apply the symmetry operation of the fibril to generate a self-interacting and fibril-interacting stack, checking that doing so will not cause a clash between the original molecule and its image. Absent a clash, we retain that pose and add the ligand-ligand van der Waals energy to the ligands docking score (here using DOCK3.8). We can check these geometries and energies using an implementation of ANI, a neural-network-based quantum-mechanical evaluation of the ligand stacking energies. In retrospective calculations, symmetry docking can reproduce the poses of three tau PET tracers whose structures have been determined. More convincingly, in a prospective study, SymDOCK predicted the structure of the PET tracer MK-6240 bound in a symmetrical stack to AD PHF tau before that structure was determined; the docked pose was used to determine how MK-6240 fit the cryo-EM density. In proof-of-concept studies, SymDOCK enriched known ligands over property-matched decoys in retrospective screens without sacrificing docking speed and can address large library screens that seek new symmetrical stackers. Future applications of this approach will be considered.
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- 2024
19. Scalable and Consolidated Microbial Platform for Rare Earth Element Leaching and Recovery from Waste Sources.
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Good, Nathan, Kang-Yun, Christina, Su, Morgan, Zytnick, Alexa, Barber, Colin, Vu, Huong, Grace, Joseph, Nguyen, Hoang, Zhang, Wenjun, Skovran, Elizabeth, Fan, Maohong, Park, Dan, and Martinez-Gomez, Norma
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acid-free leaching ,bioaccumulation ,bioconcentration ,bioleaching ,electronic waste ,lanthanide ,metal-binding protein ,neodymium ,Metals ,Rare Earth ,Metals ,Electronic Waste ,Ligands - Abstract
Chemical methods for the extraction and refinement of technologically critical rare earth elements (REEs) are energy-intensive, hazardous, and environmentally destructive. Current biobased extraction systems rely on extremophilic organisms and generate many of the same detrimental effects as chemical methodologies. The mesophilic methylotrophic bacterium Methylobacterium extorquens AM1 was previously shown to grow using electronic waste by naturally acquiring REEs to power methanol metabolism. Here we show that growth using electronic waste as a sole REE source is scalable up to 10 L with consistent metal yields without the use of harsh acids or high temperatures. The addition of organic acids increases REE leaching in a nonspecific manner. REE-specific bioleaching can be engineered through the overproduction of REE-binding ligands (called lanthanophores) and pyrroloquinoline quinone. REE bioaccumulation increases with the leachate concentration and is highly specific. REEs are stored intracellularly in polyphosphate granules, and genetic engineering to eliminate exopolyphosphatase activity increases metal accumulation, confirming the link between phosphate metabolism and biological REE use. Finally, we report the innate ability of M. extorquens to grow using other complex REE sources, including pulverized smartphones, demonstrating the flexibility and potential for use as a recovery platform for these critical metals.
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- 2024
20. An orthogonalized PYR1-based CID module with reprogrammable ligand-binding specificity.
- Author
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Park, Sang-Youl, Qiu, Jingde, Wei, Shuang, Peterson, Francis, Beltrán, Jesús, Medina-Cucurella, Angélica, Vaidya, Aditya, Xing, Zenan, Volkman, Brian, Nusinow, Dmitri, Whitehead, Timothy, Wheeldon, Ian, and Cutler, Sean
- Subjects
Abscisic Acid ,Arabidopsis ,Arabidopsis Proteins ,Dimerization ,Ligands ,Membrane Transport Proteins - Abstract
Plants sense abscisic acid (ABA) using chemical-induced dimerization (CID) modules, including the receptor PYR1 and HAB1, a phosphatase inhibited by ligand-activated PYR1. This system is unique because of the relative ease with which ligand recognition can be reprogrammed. To expand the PYR1 system, we designed an orthogonal * module, which harbors a dimer interface salt bridge; X-ray crystallographic, biochemical and in vivo analyses confirm its orthogonality. We used this module to create PYR1*MANDI/HAB1* and PYR1*AZIN/HAB1*, which possess nanomolar sensitivities to their activating ligands mandipropamid and azinphos-ethyl. Experiments in Arabidopsis thaliana and Saccharomyces cerevisiae demonstrate the sensitive detection of banned organophosphate contaminants using living biosensors and the construction of multi-input/output genetic circuits. Our new modules enable ligand-programmable multi-channel CID systems for plant and eukaryotic synthetic biology that can empower new plant-based and microbe-based sensing modalities.
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- 2024
21. Actinium chelation and crystallization in a macromolecular scaffold
- Author
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Wacker, Jennifer N, Woods, Joshua J, Rupert, Peter B, Peterson, Appie, Allaire, Marc, Lukens, Wayne W, Gaiser, Alyssa N, Minasian, Stefan G, Strong, Roland K, and Abergel, Rebecca J
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Inorganic Chemistry ,Chemical Sciences ,1.1 Normal biological development and functioning ,Actinium ,Chelating Agents ,Crystallization ,Radiopharmaceuticals ,Humans ,Ligands - Abstract
Targeted alpha therapy (TAT) pairs the specificity of antigen targeting with the lethality of alpha particles to eradicate cancerous cells. Actinium-225 [225Ac; t1/2 = 9.920(3) days] is an alpha-emitting radioisotope driving the next generation of TAT radiopharmaceuticals. Despite promising clinical results, a fundamental understanding of Ac coordination chemistry lags behind the rest of the Periodic Table due to its limited availability, lack of stable isotopes, and inadequate systems poised to probe the chemical behavior of this radionuclide. In this work, we demonstrate a platform that combines an 8-coordinate synthetic ligand and a mammalian protein to characterize the solution and solid-state behavior of the longest-lived Ac isotope, 227Ac [t1/2 = 21.772(3) years]. We expect these results to direct renewed efforts for 225Ac-TAT development, aid in understanding Ac coordination behavior relative to other +3 lanthanides and actinides, and more broadly inform this element's position on the Periodic Table.
- Published
- 2024
22. Structural characterization of ligand binding and pH-specific enzymatic activity of mouse Acidic Mammalian Chitinase
- Author
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Díaz, Roberto Efraín, Ecker, Andrew K, Correy, Galen J, Asthana, Pooja, Young, Iris D, Faust, Bryan, Thompson, Michael C, Seiple, Ian B, Van Dyken, Steven, Locksley, Richard M, and Fraser, James S
- Subjects
Biochemistry and Cell Biology ,Biological Sciences ,Underpinning research ,1.1 Normal biological development and functioning ,Chitinases ,Animals ,Hydrogen-Ion Concentration ,Mice ,Molecular Dynamics Simulation ,Chitin ,Protein Conformation ,Crystallography ,X-Ray ,Protein Binding ,Ligands ,Kinetics ,Acetylglucosamine ,Models ,Molecular ,enzyme ,chitin ,lung ,E. coli ,Human ,Mouse ,biochemistry ,chemical biology ,human ,molecular biophysics ,mouse ,structural biology ,Biological sciences ,Biomedical and clinical sciences ,Health sciences - Abstract
Chitin is an abundant biopolymer and pathogen-associated molecular pattern that stimulates a host innate immune response. Mammals express chitin-binding and chitin-degrading proteins to remove chitin from the body. One of these proteins, Acidic Mammalian Chitinase (AMCase), is an enzyme known for its ability to function under acidic conditions in the stomach but is also active in tissues with more neutral pHs, such as the lung. Here, we used a combination of biochemical, structural, and computational modeling approaches to examine how the mouse homolog (mAMCase) can act in both acidic and neutral environments. We measured kinetic properties of mAMCase activity across a broad pH range, quantifying its unusual dual activity optima at pH 2 and 7. We also solved high-resolution crystal structures of mAMCase in complex with oligomeric GlcNAcn, the building block of chitin, where we identified extensive conformational ligand heterogeneity. Leveraging these data, we conducted molecular dynamics simulations that suggest how a key catalytic residue could be protonated via distinct mechanisms in each of the two environmental pH ranges. These results integrate structural, biochemical, and computational approaches to deliver a more complete understanding of the catalytic mechanism governing mAMCase activity at different pH. Engineering proteins with tunable pH optima may provide new opportunities to develop improved enzyme variants, including AMCase, for therapeutic purposes in chitin degradation.
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- 2024
23. Optimization of cancer immunotherapy on the basis of programmed death ligand-1 distribution and function.
- Author
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Zou, Wei, Luo, Xin, Gao, Mengyuan, Yu, Chang, Wan, Xueting, Yu, Suyun, Wu, Yuanyuan, Wang, Aiyun, Wei, Zhonghong, Zhao, Yang, Lu, Yin, and Fenical, William
- Subjects
PD-L1 distribution ,PD-L1 function ,antibodies blockade ,combination therapy ,pharmacological modulators ,predictive biomarkers ,Humans ,Programmed Cell Death 1 Receptor ,B7-H1 Antigen ,Ligands ,Neoplasms ,Immunotherapy ,Tumor Microenvironment - Abstract
Programmed cell death protein-1 (PD-1)/programmed death ligand-1 (PD-L1) immune checkpoint blockade as a breakthrough in cancer immunotherapy has shown unprecedented positive outcomes in the clinic. However, the overall effectiveness of PD-L1 antibody is less than expected. An increasing number of studies have demonstrated that PD-L1 is widely distributed and expressed not only on the cell membrane but also on the inside of the cells as well as on the extracellular vesicles secreted by tumour cells. Both endogenous and exogenous PD-L1 play significant roles in influencing the therapeutic effect of anti-tumour immunity. Herein, we mainly focused on the distribution and function of PD-L1 and further summarized the potential targeted therapeutic strategies. More importantly, in addition to taking the overall expression abundance of PD-L1 as a predictive indicator for selecting corresponding PD-1/PD-L1 monoclonal antibodies (mAbs), we also proposed that personalized combination therapies based on the different distribution of PD-L1 are worth attention to achieve more efficient and effective therapeutic outcomes in cancer patients. LINKED ARTICLES: This article is part of a themed issue on Cancer Microenvironment and Pharmacological Interventions. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v181.2/issuetoc.
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- 2024
24. FGF9, a Potent Mitogen, Is a New Ligand for Integrin αvβ3, and the FGF9 Mutant Defective in Integrin Binding Acts as an Antagonist
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Chang, Chih-Chieh, Takada, Yoko K, Cheng, Chao-Wen, Maekawa, Yukina, Mori, Seiji, and Takada, Yoshikazu
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Biochemistry and Cell Biology ,Biological Sciences ,Cancer ,2.1 Biological and endogenous factors ,Aetiology ,Underpinning research ,1.1 Normal biological development and functioning ,Integrin alphaVbeta3 ,Mitogens ,Ligands ,Fibroblast Growth Factor 1 ,Fibroblast Growth Factor 2 ,DNA ,FGF9 ,integrin ,dominant-negative effect ,signaling ,FGFR3 ,mutagenesis ,docking simulation ,Biological sciences ,Biomedical and clinical sciences - Abstract
FGF9 is a potent mitogen and survival factor, but FGF9 protein levels are generally low and restricted to a few adult organs. Aberrant expression of FGF9 usually results in cancer. However, the mechanism of FGF9 action has not been fully established. Previous studies showed that FGF1 and FGF2 directly bind to integrin αvβ3, and this interaction is critical for signaling functions (FGF-integrin crosstalk). FGF1 and FGF2 mutants defective in integrin binding were defective in signaling, whereas the mutants still bound to FGFR suppressed angiogenesis and tumor growth, indicating that they act as antagonists. We hypothesize that FGF9 requires direct integrin binding for signaling. Here, we show that docking simulation of the interaction between FGF9 and αvβ3 predicted that FGF9 binds to the classical ligand-binding site of αvβ3. We show that FGF9 bound to integrin αvβ3 and generated FGF9 mutants in the predicted integrin-binding interface. An FGF9 mutant (R108E) was defective in integrin binding, activating FRS2α and ERK1/2, inducing DNA synthesis, cancer cell migration, and invasion in vitro. R108E suppressed DNA synthesis and activation of FRS2α and ERK1/2 induced by WT FGF9 (dominant-negative effect). These findings indicate that FGF9 requires direct integrin binding for signaling and that R108E has potential as an antagonist to FGF9 signaling.
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- 2024
25. Fast free energy estimates from λ-dynamics with bias-updated Gibbs sampling.
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Robo, Michael, Hayes, Ryan, Ding, Xinqiang, Pulawski, Brian, and Vilseck, Jonah
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Molecular Dynamics Simulation ,Protein Binding ,Entropy ,Thermodynamics ,Drug Design ,Ligands - Abstract
Relative binding free energy calculations have become an integral computational tool for lead optimization in structure-based drug design. Classical alchemical methods, including free energy perturbation or thermodynamic integration, compute relative free energy differences by transforming one molecule into another. However, these methods have high operational costs due to the need to perform many pairwise perturbations independently. To reduce costs and accelerate molecular design workflows, we present a method called λ-dynamics with bias-updated Gibbs sampling. This method uses dynamic biases to continuously sample between multiple ligand analogues collectively within a single simulation. We show that many relative binding free energies can be determined quickly with this approach without compromising accuracy. For five benchmark systems, agreement to experiment is high, with root mean square errors near or below 1.0 kcal mol-1. Free energy results are consistent with other computational approaches and within statistical noise of both methods (0.4 kcal mol-1 or less). Notably, large efficiency gains over thermodynamic integration of 18-66-fold for small perturbations and 100-200-fold for whole aromatic ring substitutions are observed. The rapid determination of relative binding free energies will enable larger chemical spaces to be more readily explored and structure-based drug design to be accelerated.
- Published
- 2023
26. Small Molecule Ligands of the BET-like Bromodomain, SmBRD3, Affect Schistosoma mansoni Survival, Oviposition, and Development
- Author
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Schiedel, Matthias, McArdle, Darius JB, Padalino, Gilda, Chan, Anthony KN, Forde-Thomas, Josephine, McDonough, Michael, Whiteland, Helen, Beckmann, Manfred, Cookson, Rosa, Hoffmann, Karl F, and Conway, Stuart J
- Subjects
Medicinal and Biomolecular Chemistry ,Organic Chemistry ,Chemical Sciences ,Biotechnology ,Digestive Diseases ,Infectious Diseases ,Vector-Borne Diseases ,Good Health and Well Being ,Animals ,Female ,Humans ,Schistosoma mansoni ,Oviposition ,Ligands ,Schistosomiasis ,Schistosomiasis mansoni ,Pharmacology and Pharmaceutical Sciences ,Medicinal & Biomolecular Chemistry ,Pharmacology and pharmaceutical sciences ,Medicinal and biomolecular chemistry ,Organic chemistry - Abstract
Schistosomiasis is a disease affecting >200 million people worldwide, but its treatment relies on a single agent, praziquantel. To investigate new avenues for schistosomiasis control, we have conducted the first systematic analysis of bromodomain-containing proteins (BCPs) in a causative species, Schistosoma mansoni. Having identified 29 putative bromodomains (BRDs) in 22 S. mansoni proteins, we selected SmBRD3, a tandem BRD-containing BCP that shows high similarity to the human bromodomain and extra terminal domain (BET) family, for further studies. Screening 697 small molecules identified the human BET BRD inhibitor I-BET726 as a ligand for SmBRD3. An X-ray crystal structure of I-BET726 bound to the second BRD of SmBRD3 [SmBRD3(2)] enabled rational design of a quinoline-based ligand (15) with an ITC Kd = 364 ± 26.3 nM for SmBRD3(2). The ethyl ester pro-drug of compound 15 (compound 22) shows substantial effects on sexually immature larval schistosomula, sexually mature adult worms, and snail-infective miracidia in ex vivo assays.
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- 2023
27. CD47 masks pro-phagocytic ligands in cis on tumor cells to suppress antitumor immunity
- Author
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Tang, Zhenghai, Zhong, Ming-Chao, Qian, Jin, Galindo, Cristian Camilo, Davidson, Dominique, Li, Jiaxin, Zhao, Yunlong, Hui, Enfu, and Veillette, André
- Subjects
Biomedical and Clinical Sciences ,Oncology and Carcinogenesis ,Immunology ,Cancer ,2.1 Biological and endogenous factors ,Aetiology ,Inflammatory and immune system ,Humans ,Antigens ,Differentiation ,CD47 Antigen ,Ligands ,Macrophages ,Neoplasms ,Phagocytosis ,Tumor Escape ,Animals ,Mice ,Biochemistry and cell biology - Abstract
Cancer cells often overexpress CD47, which triggers the inhibitory receptor SIRPα expressed on macrophages, to elude phagocytosis and antitumor immunity. Pharmacological blockade of CD47 or SIRPα is showing promise as anticancer therapy, although CD47 blockade has been associated with hematological toxicities that may reflect its broad expression pattern on normal cells. Here we found that, in addition to triggering SIRPα, CD47 suppressed phagocytosis by a SIRPα-independent mechanism. This mechanism prevented phagocytosis initiated by the pro-phagocytic ligand, SLAMF7, on tumor cells, due to a cis interaction between CD47 and SLAMF7. The CD47-SLAMF7 interaction was disrupted by CD47 blockade and by a first-in-class agonist SLAMF7 antibody, but not by SIRPα blockade, thereby promoting antitumor immunity. Hence, CD47 suppresses phagocytosis not only by engaging SIRPα, but also by masking cell-intrinsic pro-phagocytic ligands on tumor cells and knowledge of this mechanism may influence the decision between CD47 blockade or SIRPα blockade for therapeutic purposes.
- Published
- 2023
28. New prediction categories in CASP15
- Author
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Kryshtafovych, Andriy, Antczak, Maciej, Szachniuk, Marta, Zok, Tomasz, Kretsch, Rachael C, Rangan, Ramya, Pham, Phillip, Das, Rhiju, Robin, Xavier, Studer, Gabriel, Durairaj, Janani, Eberhardt, Jerome, Sweeney, Aaron, Topf, Maya, Schwede, Torsten, Fidelis, Krzysztof, and Moult, John
- Subjects
Biological Sciences ,Bioinformatics and Computational Biology ,Protein Conformation ,Proteins ,Models ,Molecular ,Computational Biology ,Ligands ,3D structure prediction ,CASP15 ,protein structure ,protein-ligand complexes ,RNA structure ,Mathematical Sciences ,Information and Computing Sciences ,Bioinformatics ,Biological sciences ,Mathematical sciences - Abstract
Prediction categories in the Critical Assessment of Structure Prediction (CASP) experiments change with the need to address specific problems in structure modeling. In CASP15, four new prediction categories were introduced: RNA structure, ligand-protein complexes, accuracy of oligomeric structures and their interfaces, and ensembles of alternative conformations. This paper lists technical specifications for these categories and describes their integration in the CASP data management system.
- Published
- 2023
29. ERBB Receptors and Their Ligands in the Developing Mammary Glands of Different Species: Fifteen Characters in Search of an Author
- Author
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Morato, Alessia, Accornero, Paolo, and Hovey, Russell C
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Biomedical and Clinical Sciences ,Clinical Sciences ,Cancer ,Breast Cancer ,1.1 Normal biological development and functioning ,Aetiology ,Underpinning research ,2.1 Biological and endogenous factors ,Animals ,Female ,Humans ,Mice ,Disease Models ,Animal ,ErbB Receptors ,Ligands ,Mammary Glands ,Human ,Mammary Glands ,Animal ,Hormone ,Epidermal growth factor ,Stroma ,Amphiregulin ,Oncology & Carcinogenesis ,Clinical sciences - Abstract
The ERBB tyrosine kinase receptors and their ligands belong to a complex family that has diverse biological effects and expression profiles in the developing mammary glands, where its members play an essential role in translating hormone signals into local effects. While our understanding of these processes stems mostly from mouse models, there is the potential for differences in how this family functions in the mammary glands of other species, particularly in light of their unique histomorphological features. Herein we review the postnatal distribution and function of ERBB receptors and their ligands in the mammary glands of rodents and humans, as well as for livestock and companion animals. Our analysis highlights the diverse biology for this family and its members across species, the regulation of their expression, and how their roles and functions might be modulated by varying stromal composition and hormone interactions. Given that ERBB receptors and their ligands have the potential to influence processes ranging from normal mammary development to diseased states such as cancer and/or mastitis, both in human and veterinary medicine, a more complete understanding of their biological functions should help to direct future research and the identification of new therapeutic targets.
- Published
- 2023
30. Nuclear Receptors and the Hidden Language of the Metabolome.
- Author
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Chen, Yujie, Anderson, Matthew Tom, Payne, Nathaniel, Santori, Fabio R., and Ivanova, Natalia B.
- Subjects
- *
NUCLEAR receptors (Biochemistry) , *LIGANDS (Biochemistry) , *HUMAN genome , *REGENERATIVE medicine , *TRANSCRIPTION factors - Abstract
Nuclear hormone receptors (NHRs) are a family of ligand-regulated transcription factors that control key aspects of development and physiology. The regulation of NHRs by ligands derived from metabolism or diet makes them excellent pharmacological targets, and the mechanistic understanding of how NHRs interact with their ligands to regulate downstream gene networks, along with the identification of ligands for orphan NHRs, could enable innovative approaches for cellular engineering, disease modeling and regenerative medicine. We review recent discoveries in the identification of physiologic ligands for NHRs. We propose new models of ligand-receptor co-evolution, the emergence of hormonal function and models of regulation of NHR specificity and activity via one-ligand and two-ligand models as well as feedback loops. Lastly, we discuss limitations on the processes for the identification of physiologic NHR ligands and emerging new methodologies that could be used to identify the natural ligands for the remaining 17 orphan NHRs in the human genome. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
31. Targeted Lipid-Based Drug Delivery Systems for Lung Cancer Therapy.
- Author
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Apostolou, Maria, Fatokun, Amos A., Assi, Sulaf, and Khan, Iftikhar
- Subjects
DRUG delivery systems ,LUNG cancer ,LIGANDS (Biochemistry) ,CANCER treatment ,DISEASE risk factors - Abstract
The aim of this study was to review the literature to explore the lipid-based drug delivery systems that have been investigated for improved treatment of lung cancers. Such lipid-based drug delivery systems include microemulsions, liposomes, transferosomes, niosomes, solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs). In order to minimise the side effects of chemotherapeutic active pharmaceutical ingredients, surface modification with various ligands has been introduced so that the delivery system will attach only to specific receptors which are overexpressed in lung cancer cells. This review briefly explored cancers and their aetiologies and risk factors, especially lung cancer. It then discussed different modifications that have been performed on the drug delivery systems to successfully treat lung cancer. The use of different ligands has also been investigated in this review. The particle size of drug delivery systems after the attachment of the ligand remained small, varying from 75 to 189 nm, which was the most significant physicochemical property during development as it affected the delivery of particles to specific sites in the lungs. Overall, evidence suggests that surface modified lipid-based drug delivery systems have significant potential to revolutionise the treatment of lung cancer, leading to reduced side effects from chemotherapy. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
32. LemPhos – New P-Chiral Phospholene Core Based Ligand.
- Author
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Włodarczyk, Adam and Ponikiewski, Łukasz
- Subjects
- *
PHOSPHINE oxides , *SERVER farms (Computer network management) , *ELIMINATION reactions , *IRIDIUM catalysts - Abstract
This article discusses the synthesis of a new P-chiral phospholene core-based ligand called LemPhos. The authors propose a method using a chiral diene to create diastereomers that are easier to separate, allowing for the production of P-chiral ligands. The ligands are synthesized and characterized, and their application as catalysts in asymmetric hydrogenation reactions is explored. The document also includes experimental procedures and spectroscopic data for the synthesized compounds. The research is dedicated to the memory of Stanisław Lem, a renowned science-fiction writer. [Extracted from the article]
- Published
- 2024
- Full Text
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33. Unravelling G protein‐coupled receptor signalling networks using global phosphoproteomics.
- Author
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Pokhrel, Rina, Morgan, Alexandra L., Robinson, Harley R., Stone, Martin J., and Foster, Simon R.
- Subjects
- *
G protein coupled receptors , *DRUG receptors , *PROTEOMICS , *DRUG discovery , *CELLULAR signal transduction - Abstract
G protein‐coupled receptor (GPCR) activation initiates signalling via a complex network of intracellular effectors that combine to produce diverse cellular and tissue responses. Although we have an advanced understanding of the proximal events following receptor stimulation, the molecular detail of GPCR signalling further downstream often remains obscure. Unravelling these GPCR‐mediated signalling networks has important implications for receptor biology and drug discovery. In this context, phosphoproteomics has emerged as a powerful approach for investigating global GPCR signal transduction. Here, we provide a brief overview of the phosphoproteomic workflow and discuss current limitations and future directions for this technology. By highlighting some of the novel insights into GPCR signalling networks gained using phosphoproteomics, we demonstrate the utility of global phosphoproteomics to dissect GPCR signalling networks and to accelerate discovery of new targets for therapeutic development. LINKED ARTICLES: This article is part of a themed issue Therapeutic Targeting of G Protein‐Coupled Receptors: hot topics from the Australasian Society of Clinical and Experimental Pharmacologists and Toxicologists 2021 Virtual Annual Scientific Meeting. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v181.14/issuetoc [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
34. Palladium(II)‐Catalyzed Nondirected Late‐Stage C(sp2)−H Deuteration of Heteroarenes Enabled Through a Multi‐Substrate Screening Approach.
- Author
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Dey, Jyotirmoy, Kaltenberger, Simon, and van Gemmeren, Manuel
- Subjects
- *
HETEROARENES , *DEUTERATION , *DEUTERIUM compounds , *PALLADIUM catalysts , *PALLADIUM - Abstract
The importance of deuterium labelling in a variety of applications, ranging from mechanistic studies to drug‐discovery, has spurred immense interest in the development of new methods for its efficient incorporation in organic, and especially in bioactive molecules. The five‐membered heteroarenes at the center of this work are ubiquitous motifs in bioactive molecules and efficient methods for the deuterium labelling of these compounds are therefore highly desirable. However, the profound differences in chemical properties encountered between different heteroarenes hamper the development of a single set of broadly applicable reaction conditions, often necessitating a separate optimization campaign for a given type of heteroarene. In this study we describe the use of a multi‐substrate screening approach to identify optimal reaction conditions for different classes of heteroarenes from a minimal number of screening reactions. Using this approach, four sets of complementary reaction conditions derived from our dual ligand‐based palladium catalysts for nondirected C(sp2)−H activation were identified, that together enable the deuteration of structurally diverse heteroarenes, including bioactive molecules. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
35. Azophosphines: Synthesis, Structure and Coordination Chemistry.
- Author
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Jordan, Emma J., Calder, Ethan D. E., Adcock, Holly V., Male, Louise, Nieger, Martin, Slootweg, J. Chris, and Jupp, Andrew R.
- Subjects
- *
COORDINATE covalent bond , *FUNCTIONAL groups , *TRIAZENES , *PHOSPHORUS - Abstract
The conceptual replacement of nitrogen with phosphorus in common organic functional groups unlocks new properties and reactivity. The phosphorus‐containing analogues of triazenes are underexplored but offer great potential as flexible and small bite‐angle ligands. This manuscript explores the synthesis and characterisation of a family of air‐stable azophosphine‐borane complexes, and their subsequent deprotection to the free azophosphines. These compounds are structurally characterised, both experimentally and computationally, and highlight the availability of the phosphorus lone pair for coordination. This is confirmed by demonstrating that neutral azophosphines can act as ligands in Ru complexes, and can coordinate as monodentate or bidentate ligands in a controlled manner, in contrast to their nitrogen analogues. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
36. Chemoselective Post‐Synthesis Modification of Pyridyl‐Substituted, Aromatic Phosphorus Heterocycles: Cationic Ligands for Coordination Chemistry.
- Author
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Dettling, Lea, Papke, Martin, Ernst, Moritz J., Weber, Manuela, and Müller, Christian
- Subjects
- *
LIGANDS (Chemistry) , *COORDINATE covalent bond , *COORDINATION polymers , *PHOSPHORUS , *HETEROCYCLIC compounds , *COPPER - Abstract
Triazaphospholes are potential polydentate ligands due to the presence of both phosphorus and nitrogen donor atoms within the aromatic 5‐membered heterocycle. The incorporation of an additional pyridyl‐substituent opens up the possibility of a post‐synthesis modification via chemoselective and also stepwise alkylation exclusively of the nitrogen atoms. This can be controlled by the choice and by the stoichiometry of the electrophile and allows the targeted synthesis of a variety of novel mono‐ and dicationic ligands. Reaction with Cu(I)‐halides causes the formation of cuprates of the type [CuXn](n−1)−, which enables coordination of the π‐acidic phosphorus donor to the negatively charged metal core, which is favored over the coordination by a strongly σ‐donating nitrogen atom. The use of cationic triazaphosphole derivatives can be used as a strategy to enforce the coordination of the ligand to an electron rich metal solely via the phosphorus atom. However, there is a subtle balance between the formation of either coordination polymers or dimeric structures, as the substitution pattern on the heterocycle and the nature of the halide have a large influence on the coordination motifs formed. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
37. Activation of Peroxisome Proliferator-Activated Receptor-β/δ (PPARβ/δ) in Keratinocytes by Endogenous Fatty Acids.
- Author
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Zhu, Bokai, Zhu, Xiaoyang, Borland, Michael G., Ralph, Douglas H., Chiaro, Christopher R., Krausz, Kristopher W., Ntambi, James M., Glick, Adam B., Patterson, Andrew D., Perdew, Gary H., Gonzalez, Frank J., and Peters, Jeffrey M.
- Subjects
- *
FATTY acids , *PEROXISOME proliferator-activated receptors , *NUCLEAR receptors (Biochemistry) , *MONOUNSATURATED fatty acids , *KERATINOCYTES , *ANGIOPOIETIN-like proteins , *CELL anatomy ,KERATINOCYTE differentiation - Abstract
Nuclear hormone receptors exist in dynamic equilibrium between transcriptionally active and inactive complexes dependent on interactions with ligands, proteins, and chromatin. The present studies examined the hypothesis that endogenous ligands activate peroxisome proliferator-activated receptor-β/δ (PPARβ/δ) in keratinocytes. The phorbol ester treatment or HRAS infection of primary keratinocytes increased fatty acids that were associated with enhanced PPARβ/δ activity. Fatty acids caused PPARβ/δ-dependent increases in chromatin occupancy and the expression of angiopoietin-like protein 4 (Angptl4) mRNA. Analyses demonstrated that stearoyl Co-A desaturase 1 (Scd1) mediates an increase in intracellular monounsaturated fatty acids in keratinocytes that act as PPARβ/δ ligands. The activation of PPARβ/δ with palmitoleic or oleic acid causes arrest at the G2/M phase of the cell cycle of HRAS-expressing keratinocytes that is not found in similarly treated HRAS-expressing Pparb/d-null keratinocytes. HRAS-expressing Scd1-null mouse keratinocytes exhibit enhanced cell proliferation, an effect that is mitigated by treatment with palmitoleic or oleic acid. Consistent with these findings, the ligand activation of PPARβ/δ with GW0742 or oleic acid prevented UVB-induced non-melanoma skin carcinogenesis, an effect that required PPARβ/δ. The results from these studies demonstrate that PPARβ/δ has endogenous roles in keratinocytes and can be activated by lipids found in diet and cellular components. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
38. Binding selectivity analysis of new delhi metallo-beta-lactamase-1 inhibitors using molecular dynamics simulations: Exploring possibilities for decoding antimicrobial drug resistance.
- Author
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Haque, Shafiul, Ahmad, Faraz, Mathkor, Darin Mansor, Makhdoom, Hatim, Johargy, Ayman K., Faidah, Hani, Babalghith, Ahmad O., Jalal, Naif A., Alhindi, Zain, and Bantun, Farkad
- Abstract
New Delhi metallo-beta-lactamase-1 (NDM1) confers resistance to several bacterial species against a broad range of beta-lactam antibiotics and turning them into superbugs that pose a significant threat to healthcare systems worldwide. As such, it is a potentially relevant biological target for counteracting bacterial infections. Given the lack of effective treatment options against NDM1 producing bacteria, finding a reliable inhibitor for the NDM1 enzyme is crucial. Using molecular dynamics simulations, the binding selectivities and affinities of three ligands, viz. PNK, 3S0, and N1G were investigated against NDM1. The results indicate that N1G binds with more affinity to NDM1 than PNK and 3S0. The binding energy decomposition analysis revealed that residues I35, W93, H189, K211, and N220 showed significant binding energies with PNK, 3S0, and N1G, and hence are crucially involved in the binding of the ligands to NDM1. Molecular dynamics trajectory analysis further elicited that the ligands influence dynamic flexibility of NDM1 morphology, which contributes to the partial selectivities of PNK, 3S0, and N1G. This in silico study offers a vital information for developing potential NDM1 inhibitors with high selectivity. Nevertheless, in vitro and in vivo experimental validation is mandated to extend the possible applications of these ligands as NDM1 inhibitors that succor in combating antimicrobial resistance. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
39. In Situ Structural Observation of a Substrate- and Peroxide-Bound High-Spin Ferric-Hydroperoxo Intermediate in the P450 Enzyme CYP121
- Author
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Nguyen, Romie C, Davis, Ian, Dasgupta, Medhanjali, Wang, Yifan, Simon, Philipp S, Butryn, Agata, Makita, Hiroki, Bogacz, Isabel, Dornevil, Kednerlin, Aller, Pierre, Bhowmick, Asmit, Chatterjee, Ruchira, Kim, In-Sik, Zhou, Tiankun, Mendez, Derek, Paley, Daniel W, Fuller, Franklin, Mori, Roberto Alonso, Batyuk, Alexander, Sauter, Nicholas K, Brewster, Aaron S, Orville, Allen M, Yachandra, Vittal K, Yano, Junko, Kern, Jan F, and Liu, Aimin
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Inorganic Chemistry ,Chemical Sciences ,Bioengineering ,Good Health and Well Being ,Peroxides ,Peracetic Acid ,Ligands ,Cytochrome P-450 Enzyme System ,Iron ,Heme ,Tyrosine ,Carbon ,General Chemistry ,Chemical sciences ,Engineering - Abstract
The P450 enzyme CYP121 from Mycobacterium tuberculosis catalyzes a carbon-carbon (C-C) bond coupling cyclization of the dityrosine substrate containing a diketopiperazine ring, cyclo(l-tyrosine-l-tyrosine) (cYY). An unusual high-spin (S = 5/2) ferric intermediate maximizes its population in less than 5 ms in the rapid freeze-quenching study of CYP121 during the shunt reaction with peracetic acid or hydrogen peroxide in acetic acid solution. We show that this intermediate can also be observed in the crystalline state by EPR spectroscopy. By developing an on-demand-rapid-mixing method for time-resolved serial femtosecond crystallography with X-ray free-electron laser (tr-SFX-XFEL) technology covering the millisecond time domain and without freezing, we structurally monitored the reaction in situ at room temperature. After a 200 ms peracetic acid reaction with the cocrystallized enzyme-substrate microcrystal slurry, a ferric-hydroperoxo intermediate is observed, and its structure is determined at 1.85 Å resolution. The structure shows a hydroperoxyl ligand between the heme and the native substrate, cYY. The oxygen atoms of the hydroperoxo are 2.5 and 3.2 Å from the iron ion. The end-on binding ligand adopts a near-side-on geometry and is weakly associated with the iron ion, causing the unusual high-spin state. This compound 0 intermediate, spectroscopically and structurally observed during the catalytic shunt pathway, reveals a unique binding mode that deviates from the end-on compound 0 intermediates in other heme enzymes. The hydroperoxyl ligand is only 2.9 Å from the bound cYY, suggesting an active oxidant role of the intermediate for direct substrate oxidation in the nonhydroxylation C-C bond coupling chemistry.
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- 2023
40. Mutate and Conjugate: A Method to Enable Rapid In-Cell Target Validation
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Thomas, Adam M, Serafini, Marta, Grant, Emma K, Coombs, Edward AJ, Bluck, Joseph P, Schiedel, Matthias, McDonough, Michael A, Reynolds, Jessica K, Lee, Bernadette, Platt, Michael, Sharlandjieva, Vassilena, Biggin, Philip C, Duarte, Fernanda, Milne, Thomas A, Bush, Jacob T, and Conway, Stuart J
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Medicinal and Biomolecular Chemistry ,Chemical Sciences ,Rare Diseases ,Genetics ,5.1 Pharmaceuticals ,Development of treatments and therapeutic interventions ,Generic health relevance ,Humans ,Nuclear Proteins ,Ligands ,HEK293 Cells ,Transcription Factors ,Mutant Proteins ,Cell Cycle Proteins ,Biological Sciences ,Organic Chemistry ,Biological sciences ,Chemical sciences - Abstract
Target validation remains a challenge in drug discovery, which leads to a high attrition rate in the drug discovery process, particularly in Phase II clinical trials. Consequently, new approaches to enhance target validation are valuable tools to improve the drug discovery process. Here, we report the combination of site-directed mutagenesis and electrophilic fragments to enable the rapid identification of small molecules that selectively inhibit the mutant protein. Using the bromodomain-containing protein BRD4 as an example, we employed a structure-based approach to identify the L94C mutation in the first bromodomain of BRD4 [BRD4(1)] as having a minimal effect on BRD4(1) function. We then screened a focused, KAc mimic-containing fragment set and a diverse fragment library against the mutant and wild-type proteins and identified a series of fragments that showed high selectivity for the mutant protein. These compounds were elaborated to include an alkyne click tag to enable the attachment of a fluorescent dye. These clickable compounds were then assessed in HEK293T cells, transiently expressing BRD4(1)WT or BRD4(1)L94C, to determine their selectivity for BRD4(1)L94C over other possible cellular targets. One compound was identified that shows very high selectivity for BRD4(1)L94C over all other proteins. This work provides a proof-of-concept that the combination of site-directed mutagenesis and electrophilic fragments, in a mutate and conjugate approach, can enable rapid identification of small molecule inhibitors for an appropriately mutated protein of interest. This technology can be used to assess the cellular phenotype of inhibiting the protein of interest, and the electrophilic ligand provides a starting point for noncovalent ligand development.
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- 2023
41. CCR5 drives NK cell-associated airway damage in pulmonary ischemia-reperfusion injury.
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Santos, Jesse, Wang, Ping, Shemesh, Avishai, Liu, Fengchun, Tsao, Tasha, Aguilar, Oscar, Cleary, Simon, Singer, Jonathan, Gao, Ying, Hays, Steven, Golden, Jeffrey, Leard, Lorriana, Kleinhenz, Mary, Kolaitis, Nicholas, Shah, Rupal, Venado, Aida, Weigt, S, Belperio, John, Looney, Mark, Calabrese, Daniel, Kukreja, Jasleen, Lanier, Lewis, and Greenland, John
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Chemokines ,Innate immunity ,NK cells ,Pulmonology ,Transplantation ,Animals ,Humans ,Mice ,Killer Cells ,Natural ,Ligands ,Lung ,Lung Injury ,Receptors ,CCR5 ,Reperfusion Injury - Abstract
Primary graft dysfunction (PGD) limits clinical benefit after lung transplantation, a life-prolonging therapy for patients with end-stage disease. PGD is the clinical syndrome resulting from pulmonary ischemia-reperfusion injury (IRI), driven by innate immune inflammation. We recently demonstrated a key role for NK cells in the airways of mouse models and human tissue samples of IRI. Here, we used 2 mouse models paired with human lung transplant samples to investigate the mechanisms whereby NK cells migrate to the airways to mediate lung injury. We demonstrate that chemokine receptor ligand transcripts and proteins are increased in mouse and human disease. CCR5 ligand transcripts were correlated with NK cell gene signatures independently of NK cell CCR5 ligand secretion. NK cells expressing CCR5 were increased in the lung and airways during IRI and had increased markers of tissue residency and maturation. Allosteric CCR5 drug blockade reduced the migration of NK cells to the site of injury. CCR5 blockade also blunted quantitative measures of experimental IRI. Additionally, in human lung transplant bronchoalveolar lavage samples, we found that CCR5 ligand was associated with increased patient morbidity and that the CCR5 receptor was increased in expression on human NK cells following PGD. These data support a potential mechanism for NK cell migration during lung injury and identify a plausible preventative treatment for PGD.
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- 2023
42. HER3: Toward the Prognostic Significance, Therapeutic Potential, Current Challenges, and Future Therapeutics in Different Types of Cancer.
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Majumder, Avisek
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antibody–drug conjugate (ADC) ,breast cancer ,epidermal growth factor receptor (EGFR) ,human epidermal growth factor receptor 2 (HER2) ,monoclonal antibody ,non-small cell lung cancers (NSCLC) ,small-molecule inhibitors ,structure-function analyses ,targeted therapy ,Humans ,Carcinoma ,Non-Small-Cell Lung ,Prognosis ,Ligands ,Phosphatidylinositol 3-Kinases ,Receptor ,ErbB-3 ,Lung Neoplasms - Abstract
Human epidermal growth factor receptor 3 (HER3) is the only family member of the EGRF/HER family of receptor tyrosine kinases that lacks an active kinase domain (KD), which makes it an obligate binding partner with other receptors for its oncogenic role. When HER3 is activated in a ligand-dependent (NRG1/HRG) or independent manner, it can bind to other receptors (the most potent binding partner is HER2) to regulate many biological functions (growth, survival, nutrient sensing, metabolic regulation, etc.) through the PI3K-AKT-mTOR pathway. HER3 has been found to promote tumorigenesis, tumor growth, and drug resistance in different cancer types, especially breast and non-small cell lung cancer. Given its ubiquitous expression across different solid tumors and role in oncogenesis and drug resistance, there has been a long effort to target HER3. As HER3 cannot be targeted through its KD with small-molecule kinase inhibitors via the conventional method, pharmaceutical companies have used various other approaches, including blocking either the ligand-binding domain or extracellular domain for dimerization with other receptors. The development of treatment options with anti-HER3 monoclonal antibodies, bispecific antibodies, and different combination therapies showed limited clinical efficiency for various reasons. Recent reports showed that the extracellular domain of HER3 is not required for its binding with other receptors, which raises doubt about the efforts and applicability of the development of the HER3-antibodies for treatment. Whereas HER3-directed antibody-drug conjugates showed potentiality for treatment, these drugs are still under clinical trial. The currently understood model for dimerization-induced signaling remains incomplete due to the absence of the crystal structure of HER3 signaling complexes, and many lines of evidence suggest that HER family signaling involves more than the interaction of two members. This review article will significantly expand our knowledge of HER3 signaling and shed light on developing a new generation of drugs that have fewer side effects than the current treatment regimen for these patients.
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- 2023
43. Evaluation of an Image-Derived Input Function for Kinetic Modeling of Nicotinic Acetylcholine Receptor-Binding PET Ligands in Mice.
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Zammit, Matthew, Kao, Chien-Min, Zhang, Hannah, Tsai, Hsiu-Ming, Holderman, Nathanial, Mitchell, Samuel, Tanios, Eve, Bhuiyan, Mohammed, Freifelder, Richard, Kucharski, Anna, Green, William, Mukherjee, Jogeshwar, and Chen, Chin-Tu
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2-FA85380 ,Nifene ,PET ,addiction ,kinetic modeling ,nicotine ,Mice ,Animals ,Nicotine ,Brain ,Tobacco Use Disorder ,Kinetics ,Ligands ,Positron-Emission Tomography ,Receptors ,Nicotinic - Abstract
Positron emission tomography (PET) radioligands that bind with high-affinity to α4β2-type nicotinic receptors (α4β2Rs) allow for in vivo investigations of the mechanisms underlying nicotine addiction and smoking cessation. Here, we investigate the use of an image-derived arterial input function and the cerebellum for kinetic analysis of radioligand binding in mice. Two radioligands were explored: 2-[18F]FA85380 (2-FA), displaying similar pKa and binding affinity to the smoking cessation drug varenicline (Chantix), and [18F]Nifene, displaying similar pKa and binding affinity to nicotine. Time-activity curves of the left ventricle of the heart displayed similar distribution across wild type mice, mice lacking the β2-subunit for ligand binding, and acute nicotine-treated mice, whereas reference tissue binding displayed high variation between groups. Binding potential estimated from a two-tissue compartment model fit of the data with the image-derived input function were higher than estimates from reference tissue-based estimations. Rate constants of radioligand dissociation were very slow for 2-FA and very fast for Nifene. We conclude that using an image-derived input function for kinetic modeling of nicotinic PET ligands provides suitable results compared to reference tissue-based methods and that the chemical properties of 2-FA and Nifene are suitable to study receptor response to nicotine addiction and smoking cessation therapies.
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- 2023
44. Detection of a Geminate Photoproduct of Bovine Cytochrome c Oxidase by Time-Resolved Serial Femtosecond Crystallography
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Ishigami, Izumi, Carbajo, Sergio, Zatsepin, Nadia, Hikita, Masahide, Conrad, Chelsie E, Nelson, Garrett, Coe, Jesse, Basu, Shibom, Grant, Thomas, Seaberg, Matthew H, Sierra, Raymond G, Hunter, Mark S, Fromme, Petra, Fromme, Raimund, Rousseau, Denis L, and Yeh, Syun-Ru
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Inorganic Chemistry ,Chemical Sciences ,Acquired Cognitive Impairment ,Brain Disorders ,Cattle ,Animals ,Electron Transport Complex IV ,Oxidation-Reduction ,Copper ,Ligands ,Oxygen ,Crystallography ,X-Ray ,Iron ,Water ,General Chemistry ,Chemical sciences ,Engineering - Abstract
Cytochrome c oxidase (CcO) is a large membrane-bound hemeprotein that catalyzes the reduction of dioxygen to water. Unlike classical dioxygen binding hemeproteins with a heme b group in their active sites, CcO has a unique binuclear center (BNC) composed of a copper atom (CuB) and a heme a3 iron, where O2 binds and is reduced to water. CO is a versatile O2 surrogate in ligand binding and escape reactions. Previous time-resolved spectroscopic studies of the CO complexes of bovine CcO (bCcO) revealed that photolyzing CO from the heme a3 iron leads to a metastable intermediate (CuB-CO), where CO is bound to CuB, before it escapes out of the BNC. Here, with a pump-probe based time-resolved serial femtosecond X-ray crystallography, we detected a geminate photoproduct of the bCcO-CO complex, where CO is dissociated from the heme a3 iron and moved to a temporary binding site midway between the CuB and the heme a3 iron, while the locations of the two metal centers and the conformation of Helix-X, housing the proximal histidine ligand of the heme a3 iron, remain in the CO complex state. This new structure, combined with other reported structures of bCcO, allows for a clearer definition of the ligand dissociation trajectory as well as the associated protein dynamics.
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- 2023
45. Atypical Chemokine Receptor 3 Senses CXC Chemokine Receptor 4 Activation Through GPCR Kinase Phosphorylation.
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Schafer, Christopher, Chen, Qiuyan, Tesmer, John, and Handel, Tracy
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Humans ,beta-Arrestins ,Chemokine CXCL12 ,G-Protein-Coupled Receptor Kinases ,HEK293 Cells ,Ligands ,Phosphorylation ,Protein Binding ,Receptors ,CXCR4 - Abstract
Atypical chemokine receptor 3 (ACKR3) is an arrestin-biased receptor that regulates extracellular chemokine levels through scavenging. The scavenging process restricts the availability of the chemokine agonist CXCL12 for the G protein-coupled receptor (GPCR) CXCR4 and requires phosphorylation of the ACKR3 C-terminus by GPCR kinases (GRKs). ACKR3 is phosphorylated by GRK2 and GRK5, but the mechanisms by which these kinases regulate the receptor are unresolved. Here we determined that GRK5 phosphorylation of ACKR3 results in more efficient chemokine scavenging and β-arrestin recruitment than phosphorylation by GRK2 in HEK293 cells. However, co-activation of CXCR4-enhanced ACKR3 phosphorylation by GRK2 through the liberation of Gβγ, an accessory protein required for efficient GRK2 activity. The results suggest that ACKR3 senses CXCR4 activation through a GRK2-dependent crosstalk mechanism, which enables CXCR4 to influence the efficiency of CXCL12 scavenging and β-arrestin recruitment to ACKR3. Surprisingly, we also found that despite the requirement for phosphorylation and the fact that most ligands promote β-arrestin recruitment, β-arrestins are dispensable for ACKR3 internalization and scavenging, suggesting a yet-to-be-determined function for these adapter proteins. Since ACKR3 is also a receptor for CXCL11 and opioid peptides, these data suggest that such crosstalk may also be operative in cells with CXCR3 and opioid receptor co-expression. Additionally, kinase-mediated receptor cross-regulation may be relevant to other atypical and G protein-coupled receptors that share common ligands. SIGNIFICANCE STATEMENT: The atypical receptor ACKR3 indirectly regulates CXCR4-mediated cell migration by scavenging their shared agonist CXCL12. Here, we show that scavenging and β-arrestin recruitment by ACKR3 are primarily dependent on phosphorylation by GRK5. However, we also show that CXCR4 co-activation enhances the contribution of GRK2 by liberating Gβγ. This phosphorylation crosstalk may represent a common feedback mechanism between atypical and G protein-coupled receptors with shared ligands for regulating the efficiency of scavenging or other atypical receptor functions.
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- 2023
46. Virtual Screening of Protein Data Bank via Docking Simulation Identified the Role of Integrins in Growth Factor Signaling, the Allosteric Activation of Integrins, and P-Selectin as a New Integrin Ligand.
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Fujita, Masaaki, Takada, Yoko, and Takada, Yoshikazu
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growth factor signaling ,integrins ,receptors ,P-Selectin ,Allosteric Regulation ,Ligands ,Cell Communication ,Intercellular Signaling Peptides and Proteins ,Immunologic Factors ,Cytokines ,Platelet Glycoprotein GPIIb-IIIa Complex - Abstract
Integrins were originally identified as receptors for extracellular matrix (ECM) and cell-surface molecules (e.g., VCAM-1 and ICAM-1). Later, we discovered that many soluble growth factors/cytokines bind to integrins and play a critical role in growth factor/cytokine signaling (growth factor-integrin crosstalk). We performed a virtual screening of protein data bank (PDB) using docking simulations with the integrin headpiece as a target. We showed that several growth factors (e.g., FGF1 and IGF1) induce a integrin-growth factor-cognate receptor ternary complex on the surface. Growth factor/cytokine mutants defective in integrin binding were defective in signaling functions and act as antagonists of growth factor signaling. Unexpectedly, several growth factor/cytokines activated integrins by binding to the allosteric site (site 2) in the integrin headpiece, which is distinct from the classical ligand (RGD)-binding site (site 1). Since 25-hydroxycholesterol, a major inflammatory mediator, binds to site 2, activates integrins, and induces inflammatory signaling (e.g., IL-6 and TNFα secretion), it has been proposed that site 2 is involved in inflammatory signaling. We showed that several inflammatory factors (CX3CL1, CXCL12, CCL5, sPLA2-IIA, and P-selectin) bind to site 2 and activate integrins. We propose that site 2 is involved in the pro-inflammatory action of these proteins and a potential therapeutic target. It has been well-established that platelet integrin αIIbβ3 is activated by signals from the inside of platelets induced by platelet agonists (inside-out signaling). In addition to the canonical inside-out signaling, we showed that αIIbβ3 can be allosterically activated by inflammatory cytokines/chemokines that are stored in platelet granules (e.g., CCL5, CXCL12) in the absence of inside-out signaling (e.g., soluble integrins in cell-free conditions). Thus, the allosteric activation may be involved in αIIbβ3 activation, platelet aggregation, and thrombosis. Inhibitory chemokine PF4 (CXCL4) binds to site 2 but did not activate integrins, Unexpectedly, we found that PF4/anti-PF4 complex was able to activate integrins, indicating that the anti-PF4 antibody changed the phenotype of PF4 from inhibitory to inflammatory. Since autoantibodies to PF4 are detected in vaccine-induced thrombocytopenic thrombosis (VIPP) and autoimmune diseases (e.g., SLE, and rheumatoid arthritis), we propose that this phenomenon is related to the pathogenesis of these diseases. P-selectin is known to bind exclusively to glycans (e.g., sLex) and involved in cell-cell interaction by binding to PSGL-1 (CD62P glycoprotein ligand-1). Unexpectedly, through docking simulation, we discovered that the P-selectin C-type lectin domain functions as an integrin ligand. It is interesting that no one has studied whether P-selectin binds to integrins in the last few decades. The integrin-binding site and glycan-binding site were close but distinct. Also, P-selectin lectin domain bound to site 2 and allosterically activated integrins.
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- 2023
47. Single-cell transcriptomics suggest distinct upstream drivers of IL-17A/F in hidradenitis versus psoriasis.
- Author
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Lee, Jongmi, Li, Xuan, Lee, Hyun, Kim, Katherine, Chaparala, Vasuma, Zhou, Wei, Cao, Junyue, Lowes, Michelle, Krueger, James, Murphy, William, and Kim, Jaehwan
- Subjects
Hidradenitis suppurativa ,IL-1 ,IL-17A ,IL-17F ,IL-1R1 ,IL-23 ,T cell ,dendritic cell ,fibroblast ,keratinocyte ,psoriasis ,single-cell RNA sequencing ,type 17 T cells ,Humans ,Interleukin-17 ,Interleukin-6 ,Transcriptome ,Ligands ,Interleukin-11 ,Skin ,Psoriasis ,Keratinocytes ,Hidradenitis Suppurativa - Abstract
BACKGROUND: On the basis of the mounting evidence that type 17 T (T17) cells and increased IL-17 play a key role in driving hidradenitis suppurativa (HS) lesion development, biologic agents used previously in psoriasis that block signaling of IL-17A and/or IL-17F isoforms have been repurposed to treat HS. OBJECTIVE: Our research aimed to characterize the transcriptome of HS T17 cells compared to the transcriptome of psoriasis T17 cells, along with their ligand-receptor interactions with neighborhood immune cell subsets. METHODS: Single-cell data of 12,300 cutaneous immune cells from 8 deroofing surgical HS skin samples including dermal tunnels were compared to single-cell data of psoriasis skin (19,525 cells from 11 samples) and control skin (11,920 cells from 10 samples). All single-cell data were generated by the same protocol. RESULTS: HS T17 cells expressed lower levels of IL23R and higher levels of IL1R1 and IL17F compared to psoriasis T17 cells (P < .05). HS Treg cells expressed higher levels of IL1R1 and IL17F compared to psoriasis Treg cells (P < .05). Semimature dendritic cells were the major immune cell subsets expressing IL1B in HS, and IL-1β ligand-receptor interactions between semimature dendritic cells and T17 cells were increased in HS compared to psoriasis (P < .05). HS dermal tunnel keratinocytes expressed inflammatory cytokines (IL17C, IL1A, IL1B, and IL6) that differed from the HS epidermis keratinocytes (IL36G) (P < .05). IL6, which synergizes with IL1B to maintain cytokine expression in T17 cells, was mainly expressed by fibroblasts in HS, which also expressed IL11+ inflammatory fibroblast genes (IL11, IL24, IL6, and POSTN) involved in the paracrine IL-1/IL-6 loop. CONCLUSION: The IL-1β-T17 cell cytokine axis is likely a dominant pathway in HS with HS T17 cells activated by IL-1β signaling, unlike psoriasis T17 cells, which are activated by IL-23 signaling.
- Published
- 2023
48. Interaction of CYP3A4 with caffeine: First insights into multiple substrate binding.
- Author
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Sevrioukova, Irina
- Subjects
CYP3A4 ,caffeine ,complex ,crystal structure ,cytochrome P450 ,ligand-binding protein ,spectroscopy ,Humans ,Binding Sites ,Caffeine ,Catalytic Domain ,Cytochrome P-450 CYP3A ,Ligands ,Substrate Specificity ,Protein Binding ,Allosteric Regulation ,Crystallography ,X-Ray ,Crystallization ,Demethylation ,Heme ,Hydrophobic and Hydrophilic Interactions ,Mutation - Abstract
Human cytochrome P450 3A4 (CYP3A4) is a major drug-metabolizing enzyme that shows extreme substrate promiscuity. Moreover, its large and malleable active site can simultaneously accommodate several substrate molecules of the same or different nature, which may lead to cooperative binding and allosteric behavior. Due to difficulty of crystallization of CYP3A4-substrate complexes, it remains unknown how multiple substrates can arrange in the active site. We determined crystal structures of CYP3A4 bound to three and six molecules of caffeine, a psychoactive alkaloid serving as a substrate and modulator of CYP3A4. In the ternary complex, one caffeine binds to the active site suitably for C8-hydroxylation, most preferable for CYP3A4. In the senary complex, three caffeine molecules stack parallel to the heme with the proximal ligand poised for 3-N-demethylation. However, the caffeine stack forms extensive hydrophobic interactions that could preclude product dissociation and multiple turnovers. In both complexes, caffeine is also bound in the substrate channel and on the outer surface known as a peripheral site. At all sites, aromatic stacking with the caffeine ring(s) is likely a dominant interaction, while direct and water-mediated polar contacts provide additional stabilization for the substrate-bound complexes. Protein-ligand interactions via the active site R212, intrachannel T224, and peripheral F219 were experimentally confirmed, and the latter two residues were identified as important for caffeine association. Collectively, the structural, spectral, and mutagenesis data provide valuable insights on the ligand binding mechanism and help better understand how purine-based pharmaceuticals and other aromatic compounds could interact with CYP3A4 and mediate drug-drug interactions.
- Published
- 2023
49. Binding selectivity analysis of new delhi metallo-beta-lactamase-1 inhibitors using molecular dynamics simulations: Exploring possibilities for decoding antimicrobial drug resistance
- Author
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Shafiul Haque, Faraz Ahmad, Darin Mansor Mathkor, Hatim Makhdoom, Ayman K. Johargy, Hani Faidah, Ahmad O. Babalghith, Naif A. Jalal, Zain Alhindi, and Farkad Bantun
- Subjects
New Delhi metallo-beta-lactamase 1 (NDM1) ,Molecular dynamics ,In silico ,Antibiotic resistance ,Ligands ,Infectious and parasitic diseases ,RC109-216 ,Public aspects of medicine ,RA1-1270 - Abstract
Background: New Delhi metallo-beta-lactamase-1 (NDM1) confers resistance to several bacterial species against a broad range of beta-lactam antibiotics and turning them into superbugs that pose a significant threat to healthcare systems worldwide. As such, it is a potentially relevant biological target for counteracting bacterial infections. Given the lack of effective treatment options against NDM1 producing bacteria, finding a reliable inhibitor for the NDM1 enzyme is crucial. Methods: Using molecular dynamics simulations, the binding selectivities and affinities of three ligands, viz. PNK, 3S0, and N1G were investigated against NDM1. Results: The results indicate that N1G binds with more affinity to NDM1 than PNK and 3S0. The binding energy decomposition analysis revealed that residues I35, W93, H189, K211, and N220 showed significant binding energies with PNK, 3S0, and N1G, and hence are crucially involved in the binding of the ligands to NDM1. Molecular dynamics trajectory analysis further elicited that the ligands influence dynamic flexibility of NDM1 morphology, which contributes to the partial selectivities of PNK, 3S0, and N1G. Conclusions: This in silico study offers a vital information for developing potential NDM1 inhibitors with high selectivity. Nevertheless, in vitro and in vivo experimental validation is mandated to extend the possible applications of these ligands as NDM1 inhibitors that succor in combating antimicrobial resistance.
- Published
- 2024
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50. Circulating immune checkpoints predict heart failure outcomes.
- Author
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Screever, Elles, Yousif, Laura, Moslehi, Javid, Salem, Joe-Elie, Voors, Adriaan, Silljé, Herman, de Boer, Rudolf, and Meijers, Wouter
- Subjects
Galectin-9 ,Heart failure ,Immune checkpoints ,PD-L1 ,PD-L2 ,Humans ,Animals ,Mice ,Ligands ,Heart Failure ,Prognosis ,Galectins ,Galectin 3 ,Chronic Disease - Abstract
AIMS: There are limited data examining the role of immune checkpoint (IC) ligands in the pathophysiology of heart failure (HF). Therefore, we explore this in three HF animal models and in three different human cohorts (healthy, stable, and worsening HF). METHODS AND RESULTS: Transcriptomic analyses of cardiac tissue of three different HF mouse models revealed differentially expressed IC receptors and their ligands compared with control mice. Based on this observation, serum levels of three well-known IC ligands (i.e. sPD-L1, sPD-L2 and galectin-9) were measured in stable HF patients from the Vitamin D Chronic Heart Failure (VitD-CHF) study (n = 101), as well as healthy individuals from the Prevention of Renal and Vascular End-stage Disease (PREVEND) study (n = 58). sPD-L1, sPD-L2, and galectin-9 were all associated with New York Heart Association classification. In multivariate linear regression analyses, all three IC ligands were associated with galectin-3 (β = 0.230, β = 0.283, and β = 0.304, respectively). sPD-L1 and galectin-9 were also associated with hs-troponin-T (β = 0.386 and β = 0.314). Regarding prognosis, higher serum levels of sPD-L1 and galectin-9 were significantly associated with increased risk for HF hospitalization and all-cause mortality [hazard ratio 1.69 (1.09-2.59) and hazard ratio 1.50 (1.06-2.12)]. Furthermore, the importance of IC ligands was tested in another stage of HF, namely worsening HF patients. In the worsening HF cohort (The BIOlogy Study to Tailored Treatment in Chronic Heart Failure) (n = 2032), sPD-L2 and galectin-9 were associated with New York Heart Association classification and significantly predicted outcome with an increased relative risk of 15% and 20%, after multivariable adjustment, respectively. CONCLUSIONS: IC ligands are expressed in cardiac disease models, and serum levels of IC ligands are elevated in HF patients, are associated with disease severity, and significantly predict prognosis. These data indicate a potential role for IC ligands in HF pathogenesis.
- Published
- 2023
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