165 results on '"Pau Gorostiza"'
Search Results
2. OPTOPHARMACOLOGICAL EFFECTS ON A NEW MOUSE MODEL OF COMPLETE RETINAL DEGENERATION: A GOOD TOOL FOR THERAPEUTIC ESSAYS?
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Víctor Paleo, Santiago Milla-Navarro, Pau Gorostiza, Francisco Germain, and Pedro De La Villa
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Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Published
- 2023
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3. EFFECTS OF NOVEL PHOTOSWITCHABLE DRUGS TARGETING M1 MUSCARINIC ACETYLCHOLINE RECEPTORS ON CORTICAL SLOW OSCILLATIONS AND EPILEPTIFORM DISCHARGES
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José Manuel Sánchez-Sánchez, Almudena Barbero-Castillo, Fabio Riefolo, Maria Sanchez-Vives, and Pau Gorostiza
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Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Published
- 2023
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4. Light-dependent inhibition of clathrin-mediated endocytosis in yeast unveils conserved functions of the AP2 complex
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Davia Prischich, Núria Camarero, Javier Encinar del Dedo, Maria Cambra-Pellejà, Judit Prat, Laura Nevola, Andrés Martín-Quirós, Elena Rebollo, Laura Pastor, Ernest Giralt, María Isabel Geli, and Pau Gorostiza
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Biochemistry ,Biological sciences ,Cell biology ,Molecular biology ,Natural sciences ,Science - Abstract
Summary: Clathrin-mediated endocytosis (CME) is an essential cellular process, conserved among eukaryotes. Yeast constitutes a powerful genetic model to dissect the complex endocytic machinery, yet there is a lack of specific pharmacological agents to interfere with CME in these organisms. TL2 is a light-regulated peptide inhibitor targeting the AP2-β-adaptin/β-arrestin interaction and that can photocontrol CME with high spatiotemporal precision in mammalian cells. Here, we study endocytic protein dynamics by live-cell imaging of the fluorescently tagged coat-associated protein Sla1-GFP, demonstrating that TL2 retains its inhibitory activity in S. cerevisiae spheroplasts. This is despite the β-adaptin/β-arrestin interaction not being conserved in yeast. Our data indicate that the AP2 α-adaptin is the functional target of activated TL2. We identified as interacting partners for the α-appendage, the Eps15 and epsin homologues Ede1 and Ent1. This demonstrates that endocytic cargo loading and sensing can be executed by conserved molecular interfaces, regardless of the proteins involved.
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- 2023
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5. Phosphorylation disrupts long-distance electron transport in cytochrome c
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Alexandre M. J. Gomila, Gonzalo Pérez-Mejías, Alba Nin-Hill, Alejandra Guerra-Castellano, Laura Casas-Ferrer, Sthefany Ortiz-Tescari, Antonio Díaz-Quintana, Josep Samitier, Carme Rovira, Miguel A. De la Rosa, Irene Díaz-Moreno, Pau Gorostiza, Marina I. Giannotti, and Anna Lagunas
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Science - Abstract
Electron transfer between mitochondrial cytochrome c and subunit of cytochrome bc 1 can proceed at long distance. Here the authors investigate further the mechanism and show phosphorylation regulation of the interactions between the protein partners in the electron transport chain.
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- 2022
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6. Photoswitchable type 1 muscarinic ligands for the control of cortical slow oscillations and epileptiform discharges
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Jose Manuel Sánchez-Sánchez, Fabio Riefolo, Almudena Barbero-Castillo, Rosalba Sortino, Luca Agnetta, Marta Forcella, Miquel Bosch, Michael Decker, Pau Gorostiza, and Maria Sanchez-Vives
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Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Published
- 2023
- Full Text
- View/download PDF
7. Control of Brain State Transitions with a Photoswitchable Muscarinic Agonist
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Almudena Barbero‐Castillo, Fabio Riefolo, Carlo Matera, Sara Caldas‐Martínez, Pedro Mateos‐Aparicio, Julia F. Weinert, Aida Garrido‐Charles, Enrique Claro, Maria V. Sanchez‐Vives, and Pau Gorostiza
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brain states ,light‐mediated control ,muscarinic acetylcholine receptors ,neuromodulation ,photopharmacology ,Science - Abstract
Abstract The ability to control neural activity is essential for research not only in basic neuroscience, as spatiotemporal control of activity is a fundamental experimental tool, but also in clinical neurology for therapeutic brain interventions. Transcranial‐magnetic, ultrasound, and alternating/direct current (AC/DC) stimulation are some available means of spatiotemporal controlled neuromodulation. There is also light‐mediated control, such as optogenetics, which has revolutionized neuroscience research, yet its clinical translation is hampered by the need for gene manipulation. As a drug‐based light‐mediated control, the effect of a photoswitchable muscarinic agonist (Phthalimide‐Azo‐Iper (PAI)) on a brain network is evaluated in this study. First, the conditions to manipulate M2 muscarinic receptors with light in the experimental setup are determined. Next, physiological synchronous emergent cortical activity consisting of slow oscillations—as in slow wave sleep—is transformed into a higher frequency pattern in the cerebral cortex, both in vitro and in vivo, as a consequence of PAI activation with light. These results open the way to study cholinergic neuromodulation and to control spatiotemporal patterns of activity in different brain states, their transitions, and their links to cognition and behavior. The approach can be applied to different organisms and does not require genetic manipulation, which would make it translational to humans.
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- 2021
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8. Reversible Photocontrol of Dopaminergic Transmission in Wild-Type Animals
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Carlo Matera, Pablo Calvé, Verònica Casadó-Anguera, Rosalba Sortino, Alexandre M. J. Gomila, Estefanía Moreno, Thomas Gener, Cristina Delgado-Sallent, Pau Nebot, Davide Costazza, Sara Conde-Berriozabal, Mercè Masana, Jordi Hernando, Vicent Casadó, M. Victoria Puig, and Pau Gorostiza
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azobenzene ,behavior ,brainwave ,dopamine ,GPCR ,in vivo electrophysiology ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Understanding the dopaminergic system is a priority in neurobiology and neuropharmacology. Dopamine receptors are involved in the modulation of fundamental physiological functions, and dysregulation of dopaminergic transmission is associated with major neurological disorders. However, the available tools to dissect the endogenous dopaminergic circuits have limited specificity, reversibility, resolution, or require genetic manipulation. Here, we introduce azodopa, a novel photoswitchable ligand that enables reversible spatiotemporal control of dopaminergic transmission. We demonstrate that azodopa activates D1-like receptors in vitro in a light-dependent manner. Moreover, it enables reversibly photocontrolling zebrafish motility on a timescale of seconds and allows separating the retinal component of dopaminergic neurotransmission. Azodopa increases the overall neural activity in the cortex of anesthetized mice and displays illumination-dependent activity in individual cells. Azodopa is the first photoswitchable dopamine agonist with demonstrated efficacy in wild-type animals and opens the way to remotely controlling dopaminergic neurotransmission for fundamental and therapeutic purposes.
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- 2022
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9. Rationally designed azobenzene photoswitches for efficient two-photon neuronal excitation
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Gisela Cabré, Aida Garrido-Charles, Miquel Moreno, Miquel Bosch, Montserrat Porta-de-la-Riva, Michael Krieg, Marta Gascón-Moya, Núria Camarero, Ricard Gelabert, José M. Lluch, Félix Busqué, Jordi Hernando, Pau Gorostiza, and Ramon Alibés
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Science - Abstract
Two-photon absorption capacity of azobenzene photoswitches usually comes with a reduction in thermal stability. Here, the authors developed photoswitches with high two-photon sensitivity and enhanced cis isomer thermal lifetime for the control of glutamate receptors.
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- 2019
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10. Long distance electron transfer through the aqueous solution between redox partner proteins
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Anna Lagunas, Alejandra Guerra-Castellano, Alba Nin-Hill, Irene Díaz-Moreno, Miguel A. De la Rosa, Josep Samitier, Carme Rovira, and Pau Gorostiza
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Science - Abstract
Electron transport chains rely on interactions between redox proteins, but the distance-dependence of the electron transfer rate through the solution is unknown. Here, the authors show that the current between two redox protein partners occurs at long distances and is electrochemically gated.
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- 2018
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11. Illuminating Phenylazopyridines To Photoswitch Metabotropic Glutamate Receptors: From the Flask to the Animals
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Xavier Gómez-Santacana, Silvia Pittolo, Xavier Rovira, Marc Lopez, Charleine Zussy, James A. R. Dalton, Adèle Faucherre, Chris Jopling, Jean-Philippe Pin, Francisco Ciruela, Cyril Goudet, Jesús Giraldo, Pau Gorostiza, and Amadeu Llebaria
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Chemistry ,QD1-999 - Published
- 2016
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12. Optical control of endogenous receptors and cellular excitability using targeted covalent photoswitches
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Mercè Izquierdo-Serra, Antoni Bautista-Barrufet, Ana Trapero, Aida Garrido-Charles, Ariadna Díaz-Tahoces, Nuria Camarero, Silvia Pittolo, Sergio Valbuena, Ariadna Pérez-Jiménez, Marina Gay, Alejandro García-Moll, Carles Rodríguez-Escrich, Juan Lerma, Pedro de la Villa, Eduardo Fernández, Miquel À Pericàs, Amadeu Llebaria, and Pau Gorostiza
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Science - Abstract
Biological activity can be photoswitched by light-regulated drugs, but so far only diffusible ligands have been shown to work on endogenous receptors. Here the authors develop targeted covalent photoswitches that couple to a protein target by ligand affinity, and demonstrate photocontrol of GluK1-expressing neurons.
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- 2016
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13. New GABA amides activating GABAA-receptors
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Peter Raster, Andreas Späth, Svetlana Bultakova, Pau Gorostiza, Burkhard König, and Piotr Bregestovski
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CHO-cells ,GABA ,GABA-amides ,GABA-superagonist ,patch-clamp recording ,Science ,Organic chemistry ,QD241-441 - Abstract
We have prepared a series of new and some literature-reported GABA-amides and determined their effect on the activation of GABAA-receptors expressed in CHO cells. Special attention was paid to the purification of the target compounds to remove even traces of GABA contaminations, which may arise from deprotection steps in the synthesis. GABA-amides were previously reported to be partial, full or superagonists. In our hands these compounds were not able to activate GABAA-receptor channels in whole-cell patch-clamp recordings. New GABA-amides, however, gave moderate activation responses with a clear structure–activity relationship suggesting some of these compounds as promising molecular tools for the functional analysis of GABAA-receptors.
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- 2013
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14. Light- and Redox-Dependent Force Spectroscopy Reveals that the Interaction between Plastocyanin and Plant Photosystem I Is Favored when One Partner Is Ready for Electron Transfer
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Ricardo A. Zamora, Manuel López-Ortiz, Montserrat Sales-Mateo, Chen Hu, Roberta Croce, Rinu Abraham Maniyara, Valerio Pruneri, Marina I. Giannotti, Pau Gorostiza, Biophysics Photosynthesis/Energy, and LaserLaB - Energy
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interprotein electron transfer ,plastocyanin ,Light ,Photosystem I Protein Complex ,photosystem I ,Spectrum Analysis ,General Engineering ,General Physics and Astronomy ,Water ,Electrons ,force spectroscopy ,single molecule measurements ,Electron Transport ,Cytochrome b6f Complex ,light-dependent interaction ,General Materials Science ,SDG 7 - Affordable and Clean Energy ,SDG 6 - Clean Water and Sanitation ,Oxidation-Reduction - Abstract
Photosynthesis is a fundamental process that converts photons into chemical energy, driven by large protein complexes at the thylakoid membranes of plants, cyanobacteria, and algae. In plants, water-soluble plastocyanin (Pc) is responsible for shuttling electrons between cytochrome b6f complex and the photosystem I (PSI) complex in the photosynthetic electron transport chain (PETC). For an efficient turnover, a transient complex must form between PSI and Pc in the PETC, which implies a balance between specificity and binding strength. Here, we studied the binding frequency and the unbinding force between suitably oriented plant PSI and Pc under redox control using single molecule force spectroscopy (SMFS). The binding frequency (observation of binding-unbinding events) between PSI and Pc depends on their respective redox states. The interaction between PSI and Pc is independent of the redox state of PSI when Pc is reduced, and it is disfavored in the dark (reduced P700) when Pc is oxidized. The frequency of interaction between PSI and Pc is higher when at least one of the partners is in a redox state ready for electron transfer (ET), and the post-ET situation (PSIRed-PcOx) leads to lower binding. In addition, we show that the binding of ET-ready PcRedto PSI can be regulated externally by Mg2+ions in solution.
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- 2022
15. Donor–Acceptor Stenhouse Adduct Displaying Reversible Photoswitching in Water and Neuronal Activity
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Rossella Castagna, Galyna Maleeva, Deborah Pirovano, Carlo Matera, and Pau Gorostiza
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Farmacologia ,Polymers ,phenobarbital ,barbiturates ,Biochemistry ,neuronal activity ,Neurotransmissors ,Catalysis ,GABA ,Neurologia ,Colloid and Surface Chemistry ,GABA receptor ,photopharmacology ,gamma-Aminobutyric Acid ,Pharmacology ,Neurons ,photoswitch ,DASA ,neurobiology ,Water ,Settore CHIM/06 - Chimica Organica ,Fototeràpia ,Neurotransmitters ,General Chemistry ,Phototherapy ,photochromism ,Settore CHIM/08 - Chimica Farmaceutica ,optopharmacology ,donor–acceptor stenhouse adduct ,barbital ,Neurology ,Barbital ,Settore BIO/14 - Farmacologia - Abstract
The interest in the photochromism and functional applications of donor-acceptor Stenhouse adducts (DASAs) soared in recent years owing to their outstanding advantages and flexible design. However, their low solubility and irreversible conversion in aqueous solutions hampered exploring DASAs for biology and medicine. It is notably unknown whether the barbiturate electron acceptor group retains the pharmacological activity of drugs such as phenobarbital, which targets γ-aminobutyric acid (GABA)-type A receptors (GABAARs) in the brain. Here, we have developed the model compound DASA-barbital based on a scaffold of red-switching second-generation DASAs, and we demonstrate that it is active in GABAARs and alters the neuronal firing rate in a physiological medium at neutral pH. DASA-barbital can also be reversibly photoswitched in acidic aqueous solutions using cyclodextrin, an approved ingredient of drug formulations. These findings clarify the path toward the biological applications of DASAs and to exploit the versatility displayed in polymers and materials science.
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- 2022
16. The protein matrix of plastocyanin supports long-distance charge transport with photosystem I and the copper ion regulates its spatial span and conductance
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Manuel López-Ortiz, Ricardo A. Zamora, Marina I. Giannotti, and Pau Gorostiza Langa
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Charge exchange is the fundamental process that sustains cellular respiration and photosynthesis by shuttling electrons in a cascade of electron transfer (ET) steps between redox cofactors. While intraprotein charge exchange is well characterized in protein complexes bearing multiple redox sites, interprotein processes are less understood due to the lack of suitable experimental approaches and the dynamic nature of the interactions. Proteins constrained between electrodes are known to support electron transport (ETp) through the protein matrix even without redox cofactors, as the charges housed by redox sites in ET are furnished by the electrodes in ETp configuration. However, it is unknown whether protein ETp mechanisms apply to the interprotein medium that is present in physiological conditions. Here, we study interprotein charge exchange between plant photosystem I (PSI) and its soluble redox partner plastocyanin (Pc) and address the role of the Pc copper center. Using electrochemical scanning tunnelling spectroscopy (ECSTS) current-distance and blinking measurements we respectively quantify the spatial span of charge exchange between individual Pc/PSI pairs and ETp through transient Pc/PSI complexes. Pc devoid of the redox center (Pcapo) can exchange charge with PSI and it does so at longer distances than with the copper ion (Pcholo). Conductance bursts associated to Pcapo/PSI complex formation are higher than in Pcholo/PSI. Thus, copper ions are not required for long distance ETp between PSI and Pc but regulate its spatial span and conductance. Our results suggest that the redox center that carries the charge in Pc is not necessary to exchange it in interprotein ET through the aqueous solution, and question the canonical view of tight complex binding between redox protein partners.
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- 2023
17. Ratiometric Nanothermometer Based on a Radical Excimer for In Vivo Sensing
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Davide Blasi, Nerea Gonzalez‐Pato, Xavier Rodriguez Rodriguez, Iñigo Diez‐Zabala, Sumithra Yasaswini Srinivasan, Núria Camarero, Oriol Esquivias, Mònica Roldán, Judith Guasch, Anna Laromaine, Pau Gorostiza, Jaume Veciana, and Imma Ratera
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Biomaterials ,Nanomedicine ,Cèl·lules ,Cells ,Nanomedicina ,General Materials Science ,General Chemistry ,Biotechnology - Abstract
Ratiometric fluorescent nanothermometers with near-infrared emission play an important role in in vivo sensing since they can be used as intracellular thermal sensing probes with high spatial resolution and high sensitivity, to investigate cellular functions of interest in diagnosis and therapy, where current approaches are not effective. Herein, the temperature-dependent fluorescence of organic nanoparticles is designed, synthesized, and studied based on the dual emission, generated by monomer and excimer species, of the tris(2,4,6-trichlorophenyl)methyl radical (TTM) doping organic nanoparticles (TTMd-ONPs), made of optically neutral tris(2,4,6-trichlorophenyl)methane (TTM-αH), acting as a matrix. The excimer emission intensity of TTMd-ONPs decreases with increasing temperatures whereas the monomer emission is almost independent and can be used as an internal reference. TTMd-ONPs show a great temperature sensitivity (3.4% K-1 at 328 K) and a wide temperature response at ambient conditions with excellent reversibility and high colloidal stability. In addition, TTMd-ONPs are not cytotoxic and their ratiometric outputs are unaffected by changes in the environment. Individual TTMd-ONPs are able to sense temperature changes at the nano-microscale. In vivo thermometry experiments in Caenorhabditis elegans (C. elegans) worms show that TTMd-ONPs can locally monitor internal body temperature changes with spatio-temporal resolution and high sensitivity, offering multiple applications in the biological nanothermometry field.© 2023 The Authors. Small published by Wiley-VCH GmbH.
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- 2023
18. In Vivo Applications of Photoswitchable Bioactive Compounds
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Alexandre M.J. Gomila and Pau Gorostiza
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- 2022
19. Determination of the nanoscale electrical properties of olfactory receptor hOR1A1 and their dependence on ligand binding: Towards the development of capacitance-operated odorant biosensors
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Anna Lagunas, Christine Belloir, Loïc Briand, Pau Gorostiza, and Josep Samitier
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Smell ,Volatile Organic Compounds ,Biosensors ,Odorants ,Electrochemistry ,Biomedical Engineering ,Biophysics ,Olfacte ,General Medicine ,Biosensing Techniques ,Receptors, Odorant ,Ligands ,Biotechnology - Abstract
The transduction of odorant binding into cellular signaling by olfactory receptors (ORs) is not understood and knowing its mechanism would enable developing new pharmacology and biohybrid electronic detectors of volatile organic compounds bearing high sensitivity and selectivity. The electrical characterization of ORs in bulk experiments is subject to microscopic models and assumptions. We have directly determined the nanoscale electrical properties of ORs immobilized in a fixed orientation, and their change upon odorant binding, using electrochemical scanning tunneling microscopy (EC-STM) in near-physiological conditions. Recordings of current versus time, distance, and electrochemical potential allows determining the OR impedance parameters and their dependence with odorant binding. Our results allow validating OR structural-electrostatic models and their functional activation processes, and anticipating a novel macroscopic biosensor based on ORs.
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- 2022
20. Adrenergic Modulation With Photochromic Ligands
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Pau Gorostiza, Gemma Sangüesa, Alexandre M. J. Gomila, Rebeca Diez-Alarcia, Ernest Giralt, Laura Ramírez, J. Javier Meana, B. Preda, Eduard Guasch, Davia Prischich, Carlo Matera, Montserrat Batlle, Santiago Milla-Navarro, Pedro de la Villa, and Jordi Hernando
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Azo compounds ,Adrenergic receptor ,Receptors adrenèrgics ,Mice, Nude ,Adrenergic ,Neurotransmission ,Ligands ,Catalysis ,Neurotransmissors ,Arousal ,Adrenaline receptors ,Mice ,03 medical and health sciences ,Adrenergic Agents ,0302 clinical medicine ,Biological neural network ,medicine ,Animals ,Zebrafish ,030304 developmental biology ,0303 health sciences ,Molecular Structure ,biology ,Chemistry ,Biological activity ,General Medicine ,General Chemistry ,Neurotransmitters ,biology.organism_classification ,Photochromism ,Receptors, Adrenergic ,3. Good health ,Clonidine ,Chromogenic Compounds ,Neuroscience ,030217 neurology & neurosurgery ,medicine.drug - Abstract
Altres ajuts: CERCA Programme/Generalitat de Catalunya, Fundaluce and "la Caixa" foundations (ID 100010434, agreement LCF/PR/HR19/52160010), Co-financed by the European Union Regional Development Fund within the framework of the ERDF Operational Program of Catalonia 2014-2020 Adrenoceptors are ubiquitous and mediate important autonomic functions as well as modulating arousal, cognition, and pain on a central level. Understanding these physiological processes and their underlying neural circuits requires manipulating adrenergic neurotransmission with high spatio-temporal precision. Here we present a first generation of photochromic ligands (adrenoswitches) obtained via azologization of a class of cyclic amidines related to the known ligand clonidine. Their pharmacology, photochromism, bioavailability, and lack of toxicity allow for broad biological applications, as demonstrated by controlling locomotion in zebrafish and pupillary responses in mice.
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- 2021
21. A donor-acceptor Stenhouse adduct displaying reversible photoswitching in water and neuronal activity
- Author
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Rossella Castagna, Galyna Maleeva, Deborah Pirovano, Carlo Matera, and Pau Gorostiza
- Abstract
The interest in the photochromism and functional applications of donor-acceptor Stenhouse adducts (DASAs) soared in recent years, owing to their outstanding advantages and flexible design. However, their low solubility and irreversible conversion in aqueous solutions hampered exploring DASAs for biology and medicine. It is notably unknown whether the barbiturate electron acceptor group retains the pharmacological activity of drugs like phenobarbital, which targets γ-aminobutyric acid (GABA) type A receptors (GABAARs) in the brain. Here, we have developed the model compound DASA-barbital based on a scaffold of red-switching second-generation DASAs and we demonstrate that it is active in GABAARs and alters the neuronal firing rate in a physiological medium at neutral pH. DASA-barbital can also be reversibly photoswitched in acidic aqueous solutions using cyclodextrin, an approved ingredient of drug formulations. These findings clear the path towards the biological applications of DASAs and to exploit the versatility displayed in polymers and materials science.
- Published
- 2022
22. Determination of the Nanoscale Electrical Properties of Olfactory Receptor hOR1A1: Towards the Development of Highly Sensitive Capacitance-Operated Odorant Biosensors
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Anna Lagunas, Christine Belloir, Loïc Briand, Pau Gorostiza, and Josep Samitier
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History ,Polymers and Plastics ,Business and International Management ,Industrial and Manufacturing Engineering - Published
- 2022
23. Odorant binding changes the electrical properties of olfactory receptors at the nanoscale
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Anna Lagunas, Loïc Briand, Josep Samitier, Pau Gorostiza, Christine Belloir, Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Instituto de Salud Carlos III [Madrid] (ISC)-ministerio de ciencia e innovacion, Institute for Bioengineering of Catalonia [Barcelona] (IBEC), Centre des Sciences du Goût et de l'Alimentation [Dijon] (CSGA), Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Bourgogne Franche-Comté [COMUE] (UBFC), Institució Catalana de Recerca i Estudis Avançats (ICREA), Universitat de Barcelona (UB), The Biomedical Research Networking Center (CIBER), Spain. CIBER is an initiative funded by the VI National R&D&i Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions, and the Instituto de Salud Carlos III (RD16/0006/0012, RD16/0011/0022), with the support of the European Regional Development Fund (ERDF). This work was funded by the CERCA Program and by the Commission for Universities and Research of the Department of Innovation, Universities, and Enterprise of the Generalitat de Catalunya (2017-SGR-1079, 2017-SGR-1442 and 2017-SGR-00465). This work was supported by grants from the Conseil Régional Bourgogne Franche-Comté (PARI grant) and the FEDER (European Funding for Regional Economical Development). This research also received funding from the European Union Research and Innovation Programme Horizon 2020 (Human Brain Project SGA3 No. 945539), DEEPER (ICT-36-2020-101016787), Agency for Management of University and Research Grants of the Government of Catalonia (CERCA Programme, 2017-SGR-1442 project, AGAUR, Clúster Emergent del Cervell Humà), and Ministry of Economy and Competitiveness (Grant PID2019-111493RB-I00)., and Julien, Sabine
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Cell signaling ,Olfactory receptor ,Odorant binding ,Chemistry ,olfactory receptor ,odorant binding ,Impedance parameters ,law.invention ,[SDV.AEN] Life Sciences [q-bio]/Food and Nutrition ,medicine.anatomical_structure ,open-circuit voltage ,law ,electrochemical scanning tunneling microscopy (EC-STM) ,impedance ,[SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology ,medicine ,Biophysics ,Scanning tunneling microscope ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM] ,Receptor ,Transduction (physiology) ,[SDV.AEN]Life Sciences [q-bio]/Food and Nutrition ,Electrochemical potential - Abstract
Olfactory receptors (ORs) comprise the largest multigene family in the vertebrates. They belong to the class A (rhodopsin-like) family of G protein-coupled receptors (GPCRs), which are the most abundant membrane proteins, having widespread, significant roles in signal transduction in cells, and therefore, they are a major pharmacological target. Moreover, ORs displayed high selectivity and sensitivity towards odorant detection, a characteristic that raised the interest for developing biohybrid sensors based on ORs for the detection of volatile compounds. The transduction of odorant binding into cellular signaling by ORs is not well understood and knowing its mechanism would enable developing new pharmacology and high performance biohybrid electronic sensors. Recent findings suggest that ligand recognition by ORs is determined by the nanoscale alterations of charge distribution in the receptor structure (Ref). However, the electrical characterization of ORs and their response towards ligand binding in bulk experiments is subjected to microscopic models and assumptions [2]. Here, we have directly determined the nanoscale electrical properties of ORs with unprecedented control over the receptor orientation, and their change upon odorant binding, using electrochemical scanning tunneling microscopy (EC-STM) in near-physiological conditions. Recordings of current versus time, distance, and electrochemical potential allows determining the OR impedance parameters and their dependence with odorant binding. The simultaneous measurement of RC equivalent by means of the open-circuit voltage (VOC) allows increasing the electrical sensitivity at the single receptor level for biosensing applications. Our results allow validating OR structural-electrostatic models and their functional activation processes.
- Published
- 2021
24. PO015 / #913 REMOTE CONTROL OF BRAIN WAVES WITH LIGHT AND A PHOTOSWITCHABLE MUSCARINIC AGONIST
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Almudena Barbero-Castillo, Fabio Riefolo, Carlo Matera, Maria Victoria Sanchez-Vives, and Pau Gorostiza
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Anesthesiology and Pain Medicine ,Neurology ,Neurology (clinical) ,General Medicine - Published
- 2022
25. Control of Brain State Transitions with a Photoswitchable Muscarinic Agonist
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Sara Caldas-Martinez, Almudena Barbero-Castillo, Julia F. Weinert, Maria V. Sanchez-Vives, Enrique Claro, Carlo Matera, Pau Gorostiza, Fabio Riefolo, Pedro Mateos‐Aparicio, and Aida Garrido-Charles
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Science ,General Chemical Engineering ,General Physics and Astronomy ,Medicine (miscellaneous) ,Stimulation ,02 engineering and technology ,Biology ,Optogenetics ,Muscarinic Agonists ,010402 general chemistry ,01 natural sciences ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,Muscarinic agonist ,Neurologia ,Mice ,brain states ,In vivo ,Muscarinic acetylcholine receptor ,muscarinic acetylcholine receptors ,medicine ,Neurociències ,Animals ,General Materials Science ,photopharmacology ,Research Articles ,General Engineering ,Neurosciences ,Ferrets ,Brain ,021001 nanoscience & nanotechnology ,Neuromodulation (medicine) ,0104 chemical sciences ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Neurology ,Cerebral cortex ,light‐mediated control ,neuromodulation ,Models, Animal ,Cholinergic ,0210 nano-technology ,Neuroscience ,Research Article - Abstract
The ability to control neural activity is essential for research not only in basic neuroscience, as spatiotemporal control of activity is a fundamental experimental tool, but also in clinical neurology for therapeutic brain interventions. Transcranial‐magnetic, ultrasound, and alternating/direct current (AC/DC) stimulation are some available means of spatiotemporal controlled neuromodulation. There is also light‐mediated control, such as optogenetics, which has revolutionized neuroscience research, yet its clinical translation is hampered by the need for gene manipulation. As a drug‐based light‐mediated control, the effect of a photoswitchable muscarinic agonist (Phthalimide‐Azo‐Iper (PAI)) on a brain network is evaluated in this study. First, the conditions to manipulate M2 muscarinic receptors with light in the experimental setup are determined. Next, physiological synchronous emergent cortical activity consisting of slow oscillations—as in slow wave sleep—is transformed into a higher frequency pattern in the cerebral cortex, both in vitro and in vivo, as a consequence of PAI activation with light. These results open the way to study cholinergic neuromodulation and to control spatiotemporal patterns of activity in different brain states, their transitions, and their links to cognition and behavior. The approach can be applied to different organisms and does not require genetic manipulation, which would make it translational to humans., Brain pathologies often require drug treatments, however drugs act all over the central nervous system. Wouldn't it be good to determine where/when a drug should be active? Drugs can be made sensitive to light, to be activated at specific times and locations. This study demonstrates that a light‐activated cholinergic drug can effectively modulate activity in the cerebral cortex network.
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- 2021
- Full Text
- View/download PDF
26. Distance and Potential Dependence of Charge Transport Through the P700 Reaction Center of Photosystem I
- Author
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Pau Gorostiza, Roberta Croce, Chen Hu, Marina Inés Giannotti, Ricardo A Zamora, and Manuel López Ortiz
- Abstract
Photoinduced charge separation and transport through photosystem I (PSI) is an essential part of the photosynthetic electron transport chain. To investigate charge exchange processes mediated by the P700 reaction center of PSI, we have developed a strategy to functionalize gold electrodes with PSI complexes that orients and exposes their luminal side to the electrolyte. Bulk photoelectrochemical measurements demonstrate that PSI remains functional in a wide sample potential range around 0 mV/SSC. Electrochemical scanning tunneling microscopy (ECSTM) imaging of individual complexes shows lateral sizes in agreement with the dimensions of PSI and an apparent height that is gated by the probe potential of ECTSM as reported for smaller globular redox proteins. This experimental setup enables ECSTM current-distance spectroscopic measurements that unequivocally correspond to the P700 side of PSI. In these conditions, we observe that the spatial span of the current is enhanced (the distance-decay rate β is reduced) through the solution at sample potential 0 mV/SSC and probe potential 400 mV/SSC. This process corresponds to hole injection into an electronic state that is available in the absence of illumination. We propose that a pair of tryptophan residues located near P700 and known to integrate the hydrophobic recognition site for plastocyanin may have an additional role as hole exchange mediator involved in charge transport through PSI.
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- 2021
27. Distance and Potential Dependence of Charge Transport Through the P700 Reaction Center of Photosystem I
- Author
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Marina I. Giannotti, Manuel López Ortiz, Manuel López-Ortiz, Chen Hu, Pau Gorostiza, Ricardo A. Zamora, and Roberta Croce
- Subjects
Photosynthetic reaction centre ,P700 ,Materials science ,Photoinduced charge separation ,Chemical physics ,law ,Electrolyte ,Scanning tunneling microscope ,Electrochemistry ,Photosystem I ,Plastocyanin ,law.invention - Abstract
Photoinduced charge separation and transport through photosystem I (PSI) is an essential part of the photosynthetic electron transport chain. To investigate charge exchange processes mediated by the P700 reaction center of PSI, we have developed a strategy to functionalize gold electrodes with PSI complexes that orients and exposes their luminal side to the electrolyte. Bulk photoelectrochemical measurements demonstrate that PSI remains functional in a wide sample potential range around 0 mV/SSC. Electrochemical scanning tunneling microscopy (ECSTM) imaging of individual complexes shows lateral sizes in agreement with the dimensions of PSI and an apparent height that is gated by the probe potential of ECTSM as reported for smaller globular redox proteins. This experimental setup enables ECSTM current-distance spectroscopic measurements that unequivocally correspond to the P700 side of PSI. In these conditions, we observe that the spatial span of the current is enhanced (the distance-decay rate β is reduced) through the solution at sample potential 0 mV/SSC and probe potential 400 mV/SSC. This process corresponds to hole injection into an electronic state that is available in the absence of illumination. We propose that a pair of tryptophan residues located near P700 and known to integrate the hydrophobic recognition site for plastocyanin may have an additional role as hole exchange mediator involved in charge transport through PSI.
- Published
- 2021
28. Light-dependent inhibition of clathrin-mediated endocytosis in yeast
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Prat J, Ernest Giralt, Andrés Martín-Quirós, Isabel Geli M, Cambra M, D. Prischich, Núria Camarero, Elena Rebollo, Laura Nevola, Pau Gorostiza, and del Dedo Je
- Subjects
Chemistry ,Protein dynamics ,education ,Endocytic cycle ,Genetic model ,Receptor-mediated endocytosis ,Spheroplast ,Inhibitory postsynaptic potential ,Endocytosis ,Yeast ,Cell biology - Abstract
Clathrin-mediated endocytosis (CME) is an essential cellular process, which is evolutionarily conserved among eukaryotes. Yeast constitutes a powerful genetic model to dissect the complex endocytic machinery, yet there is a lack of pharmacological agents that could complement genetics in selectively and reversibly interfere with CME in these organisms. TL2 is a light-regulated peptide inhibitor that targets the AP2/β-arrestin interaction and that can photocontrol CME with high spatiotemporal precision in mammalian cells. Here, we study endocytic protein dynamics by live-cell imaging of the fluorescently tagged coat-associated protein Sla1-GFP and demonstrate that TL2 retains its inhibitory activity in S. cerevisiae spheroplasts, thus providing a unique tool for acute and reversible CME modulation in yeast.
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- 2021
29. Electrochemically Gated Long-Distance Charge Transport in Photosystem I
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Maria Elena Antinori, Pau Gorostiza, Ismael Díez-Pérez, Carme Rovira, Emilie Wientjes, Montserrat López-Martínez, Alba Nin-Hill, Roberta Croce, Manuel López-Ortiz, Biophysics Photosynthesis/Energy, and LaserLaB - Energy
- Subjects
Materials science ,Biophysics ,Electrochemistry ,Photosystem I ,010402 general chemistry ,Redox ,01 natural sciences ,7. Clean energy ,Catalysis ,electrochemical gating ,law.invention ,symbols.namesake ,Electron transfer ,03 medical and health sciences ,law ,Life Science ,SDG 7 - Affordable and Clean Energy ,030304 developmental biology ,0303 health sciences ,P700 ,photosynthesis ,010405 organic chemistry ,Fermi level ,General Chemistry ,General Medicine ,current decay ,electron transfer ,Electron transport chain ,0104 chemical sciences ,Biofysica ,Chemical physics ,symbols ,scanning tunneling microscopy ,Scanning tunneling microscope ,EPS - Abstract
The transport of electrons along photosynthetic and respiratory chains involves a series of enzymatic reactions that are coupled through redox mediators, including proteins and small molecules. The use of native and synthetic redox probes is key to understanding charge transport mechanisms and to the design of bioelectronic sensors and solar energy conversion devices. However, redox probes have limited tunability to exchange charge at the desired electrochemical potentials (energy levels) and at different protein sites. Herein, we take advantage of electrochemical scanning tunneling microscopy (ECSTM) to control the Fermi level and nanometric position of the ECSTM probe in order to study electron transport in individual photosystem I (PSI) complexes. Current–distance measurements at different potentiostatic conditions indicate that PSI supports long-distance transport that is electrochemically gated near the redox potential of P700, with current extending farther under hole injection conditions.
- Published
- 2019
30. Fast Photo-Chrono-Amperometry of Photosynthetic Complexes for Biosensors and Electron Transport Studies
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Maria Elena Antinori, Pau Gorostiza, Roberta Croce, Niek F. van Hulst, Chen Hu, Manuel López-Ortiz, Vikas Remesh, Ricardo A. Zamora, Biophysics Photosynthesis/Energy, and LaserLaB - Energy
- Subjects
Paraquat ,Photosystem II ,Plastoquinone ,Bioengineering ,02 engineering and technology ,Biosensing Techniques ,Photochemistry ,Photosystem I ,Photosynthesis ,01 natural sciences ,Redox ,Electron Transport ,chemistry.chemical_compound ,medicine ,Instrumentation ,Fluid Flow and Transfer Processes ,Física [Àrees temàtiques de la UPC] ,Photosystem I Protein Complex ,Process Chemistry and Technology ,010401 analytical chemistry ,Photosystem II Protein Complex ,Viologen ,021001 nanoscience & nanotechnology ,Electron transport chain ,0104 chemical sciences ,Biosensors ,chemistry ,0210 nano-technology ,Biosensor ,medicine.drug - Abstract
Photosynthetic reactions in plants, algae, and cyanobacteria are driven by photosystem I and photosystem II complexes, which specifically reduce or oxidize partner redox biomolecules. Photosynthetic complexes can also bind synthetic organic molecules, which inhibit their photoactivity and can be used both to study the electron transport chain and as herbicides and algicides. Thus, their development, characterization, and sensing bears fundamental and applied interest. Substantial efforts have been devoted to developing photosensors based on photosystem II to detect compounds that bind to the plastoquinone sites of this complex. In comparison, photosystem I based sensors have received less attention and could be used to identify novel substances displaying phytotoxic effects, including those obtained from natural product extracts. We have developed a robust procedure to functionalize gold electrodes with photo- and redox-active photosystem I complexes based on transparent gold and a thiolate self-assembled monolayer, and we have obtained reproducible electrochemical photoresponses. Chronoamperometric recordings have allowed us to measure photocurrents in the presence of the viologen derivative paraquat at concentrations below 100 nM under lock-in operation and a sensor dynamic range spanning six orders of magnitude up to 100 mM. We have modeled their time course to identify the main electrochemical processes and limiting steps in the electron transport chain. Our results allow us to isolate the contributions from photosystem I and the redox mediator, and evaluate photocurrent features (spectral and power dependence, fast transient kinetics) that could be used as a sensing signal to detect other inhibitors and modulators of photosystem I activity.
- Published
- 2021
31. Subunit-specific photocontrol of glycine receptors by azobenzene-nitrazepam photoswitcher
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Piotr Bregestovski, Daniel Wutz, Mercedes Alfonso-Prieto, Alexandre M. J. Gomila, Elena Petukhova, Daria Ponomareva, Alba Nin-Hill, Elvira Mukhametova, Galyna Maleeva, Burkhard König, Pau Gorostiza, and Karin Rustler
- Subjects
Farmacologia ,hypoglossal motoneurons ,Patch-Clamp Techniques ,Neuronal Excitability ,Drug action ,Inhibitory postsynaptic potential ,Mice ,Receptors, Glycine ,medicine ,Animals ,photopharmacology ,Channel blocker ,ddc:610 ,Hyperekplexia ,Nitrazepam ,Receptor ,Glycine receptor ,Pharmacology ,GABAA receptor ,Chemistry ,glycine receptors ,General Neuroscience ,General Medicine ,patch-clamp ,brain slices ,molecular modelling ,Transmembrane domain ,Amino acids ,Aminoàcids ,medicine.symptom ,Azo Compounds ,Neuroscience ,Research Article: New Research - Abstract
Visual Abstract, Photopharmacology is a unique approach that through a combination of photochemistry methods and advanced life science techniques allows the study and control of specific biological processes, ranging from intracellular pathways to brain circuits. Recently, a first photochromic channel blocker of anion-selective GABAA receptors, the azobenzene-nitrazepam-based photochromic compound (Azo-NZ1), has been described. In the present study, using patch-clamp technique in heterologous system and in mice brain slices, site-directed mutagenesis and molecular modeling we provide evidence of the interaction of Azo-NZ1 with glycine receptors (GlyRs) and determine the molecular basis of this interaction. Glycinergic synaptic neurotransmission determines an important inhibitory drive in the vertebrate nervous system and plays a crucial role in the control of neuronal circuits in the spinal cord and brain stem. GlyRs are involved in locomotion, pain sensation, breathing, and auditory function, as well as in the development of such disorders as hyperekplexia, epilepsy, and autism. Here, we demonstrate that Azo-NZ1 blocks in a UV-dependent manner the activity of α2 GlyRs (GlyR2), while being barely active on α1 GlyRs (GlyR1). The site of Azo-NZ1 action is in the chloride-selective pore of GlyR at the 2’ position of transmembrane helix 2 and amino acids forming this site determine the difference in Azo-NZ1 blocking activity between GlyR2 and GlyR1. This subunit-specific modulation is also shown on motoneurons of brainstem slices from neonatal mice that switch during development from expressing “fetal” GlyR2 to “adult” GlyR1 receptors.
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- 2021
32. Rationally designed azobenzene photoswitches for efficient two-photon neuronal excitation
- Author
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Félix Busqué, José M. Lluch, Núria Camarero, Miquel Bosch, Ricard Gelabert, Jordi Hernando, Gisela Cabré, Marta Gascón-Moya, Miquel Moreno, Aida Garrido-Charles, Montserrat Porta-de-la-Riva, Pau Gorostiza, Ramon Alibés, and Michael Krieg
- Subjects
0301 basic medicine ,Patch-Clamp Techniques ,Photoisomerization ,Canals de calci ,General Physics and Astronomy ,Fotobiología ,Neurones ,02 engineering and technology ,chemistry.chemical_compound ,Two-photon excitation microscopy ,Premovement neuronal activity ,neuronal excitation ,lcsh:Science ,Neurons ,Multidisciplinary ,021001 nanoscience & nanotechnology ,Photochemical Processes ,Photobiology ,Fotobiologia ,Azobenzene ,0210 nano-technology ,Materials science ,Infrared Rays ,Science ,Herramientas químicas ,Neuronal excitation ,General Biochemistry, Genetics and Molecular Biology ,Article ,Cell Line ,03 medical and health sciences ,Animals ,Humans ,Caenorhabditis elegans ,Eines químiques ,Photons ,Física [Àrees temàtiques de la UPC] ,Absorption cross section ,Rational design ,Computational Biology ,General Chemistry ,Canales de calcio ,030104 developmental biology ,HEK293 Cells ,chemistry ,Biophysics ,lcsh:Q ,Calcium Channels ,Chemical tools ,Azo Compounds ,Excitation - Abstract
Manipulation of neuronal activity using two-photon excitation of azobenzene photoswitches with near-infrared light has been recently demonstrated, but their practical use in neuronal tissue to photostimulate individual neurons with three-dimensional precision has been hampered by firstly, the low efficacy and reliability of NIR-induced azobenzene photoisomerization compared to one-photon excitation, and secondly, the short cis state lifetime of the two-photon responsive azo switches. Here we report the rational design based on theoretical calculations and the synthesis of azobenzene photoswitches endowed with both high two-photon absorption cross section and slow thermal back-isomerization. These compounds provide optimized and sustained two-photon neuronal stimulation both in light-scattering brain tissue and in Caenorhabditis elegans nematodes, displaying photoresponse intensities that are comparable to those achieved under one-photon excitation. This finding opens the way to use both genetically targeted and pharmacologically selective azobenzene photoswitches to dissect intact neuronal circuits in three dimensions., Two-photon absorption capacity of azobenzene photoswitches usually comes with a reduction in thermal stability. Here, the authors developed photoswitches with high two-photon sensitivity and enhanced cis isomer thermal lifetime for the control of glutamate receptors.
- Published
- 2021
33. Photocontrol of Endogenous Glycine Receptors In Vivo
- Author
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Elvira Mukhametova, Alba Nin-Hill, Franck Peiretti, Burkhard König, Daniel Wutz, Piotr Bregestovski, Marat A. Mukhamedyarov, Xavier Rovira, Miquel Bosch, Daria Ponomareva, Carme Rovira, Pau Gorostiza, Karin Rustler, Galyna Maleeva, Antoni Bautista-Barrufet, Alexandre M. J. Gomila, Elena Petukhova, Mercedes Alfonso-Prieto, Institute for Bioengineering of Catalonia [Barcelona] (IBEC), Barcelona Institute of Science and Technology (BIST), University of Regensburg, Institut de Neurosciences des Systèmes (INS), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Química Teòrica i Computacional (IQTCUB), Universitat de Barcelona (UB), University of Barcelona, Kazan Federal University (KFU), Kazan State University (KPFU), Centre recherche en CardioVasculaire et Nutrition = Center for CardioVascular and Nutrition research (C2VN), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institute of Neuroscience and Medicine [Jülich] (INM-1), Heinrich Heine Universität Düsseldorf = Heinrich Heine University [Düsseldorf], Universitat de Barcelona and the Intitut de Recerca en Química Teòrica i Computacional (IQTCUB), Institució Catalana de Recerca i Estudis Avançats (ICREA), ERA SynBIO grant MODULIGHTOR PCIN-2015-163-C02-01Russian Science Foundation (RSF) 18-15-00313AGAUR/Generalitat de Catalunya (CERCA Programme) 2017-SGR-1442Human Brain Project WAVESCALES (SGA2 Grant) 785907MINECO CTQ2016-80066R, and peiretti, franck
- Subjects
Male ,Clinical Biochemistry ,Allosteric regulation ,Neurotransmission ,Biology ,Inhibitory postsynaptic potential ,Ligands ,01 natural sciences ,Biochemistry ,Synaptic Transmission ,Mice ,[SCCO]Cognitive science ,Cricetulus ,Receptors, Glycine ,In vivo ,ddc:570 ,Drug Discovery ,medicine ,Animals ,photopharmacology ,Receptor ,Molecular Biology ,Glycine receptor ,Cells, Cultured ,Pharmacology ,Neurons ,Mice, Inbred ICR ,Molecular Structure ,010405 organic chemistry ,glycine receptors ,[SCCO] Cognitive science ,Photochemical Processes ,3. Good health ,0104 chemical sciences ,optopharmacology ,Molecular Docking Simulation ,inhibitory neurotransmission ,in vivo ,Mechanism of action ,Excitatory postsynaptic potential ,Molecular Medicine ,Female ,photoswitch ,medicine.symptom ,CNS ,Neuroscience ,Azo Compounds - Abstract
International audience; Glycine receptors (GlyRs) are indispensable for maintaining excitatory/inhibitory balance in neuronal circuits that control reflexes and rhythmic motor behaviors. Here we have developed Glyght, a GlyR ligand controlled with light. It is selective over other Cys-loop receptors, is active in vivo, and displays an allosteric mechanism of action. The photomanipulation of glycinergic neurotransmission opens new avenues to understanding inhibitory circuits in intact animals and to developing drug-based phototherapies.
- Published
- 2020
34. Optical Control of GABA
- Author
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Karin, Rustler, Galyna, Maleeva, Alexandre M J, Gomila, Pau, Gorostiza, Piotr, Bregestovski, and Burkhard, König
- Subjects
Benzodiazepines ,in vivo ,fulgimides ,GABAA Receptors ,Communication ,photopharmacology ,Receptors, GABA-A ,Biochemistry ,gamma-Aminobutyric Acid ,Communications - Abstract
Optogenetic and photopharmacological tools to manipulate neuronal inhibition have limited efficacy and reversibility. We report the design, synthesis, and biological evaluation of Fulgazepam, a fulgimide derivative of benzodiazepine that behaves as a pure potentiator of ionotropic γ‐aminobutyric acid receptors (GABAARs) and displays full and reversible photoswitching in vitro and in vivo. The compound enables high‐resolution studies of GABAergic neurotransmission, and phototherapies based on localized, acute, and reversible neuroinhibition., Fulgazepam: Herein, the design, synthesis, and biological evaluation of Fulgazepam, a fulgimide derivative of benzodiazepine that behaves as a pure potentiator of ionotropic γ‐aminobutyric acid receptors (GABAARs) and displays full and reversible photoswitching in vitro and in vivo, is reported.
- Published
- 2020
35. Kainate Receptor Activation Shapes Short-Term Synaptic Plasticity by Controlling Receptor Lateral Mobility at Glutamatergic Synapses
- Author
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Thierry Nieus, Alice Polenghi, Stefania Guazzi, Pau Gorostiza, Enrica Maria Petrini, and Andrea Barberis
- Subjects
0301 basic medicine ,Postsynaptic Current ,Glutamic Acid ,catenin ,Kainate receptor ,Optogenetics ,Synaptic Transmission ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,Glutamatergic ,0302 clinical medicine ,glutamatergic synapses ,Postsynaptic potential ,lateral diffusion ,Humans ,Receptors neurals ,glutamate uncaging ,Neural transmission ,Kainic Acid ,Neuronal Plasticity ,Chemistry ,Glutamate binding ,short-term synaptic plasticity ,glutamatergic PSD ,Crosstalk (biology) ,030104 developmental biology ,cadherin ,Sinapsi ,Synaptic plasticity ,Synapses ,Neurotransmissió ,single-particle tracking ,kainate receptors ,model simulations ,Neuroscience ,Neural receptor ,030217 neurology & neurosurgery - Abstract
Summary Kainate receptors (KARs) mediate postsynaptic currents with a key impact on neuronal excitability. However, the molecular determinants controlling KAR postsynaptic localization and stabilization are poorly understood. Here, we exploit optogenetic and single-particle tracking approaches to study the role of KAR conformational states induced by glutamate binding on KAR lateral mobility at synapses. We report that following glutamate binding, KARs are readily and reversibly trapped at glutamatergic synapses through increased interaction with the β-catenin/N-cadherin complex. We demonstrate that such activation-dependent synaptic immobilization of KARs is crucial for the modulation of short-term plasticity of glutamatergic synapses. Thus, the present study unveils the crosstalk between conformational states and lateral mobility of KARs, a mechanism regulating glutamatergic signaling, particularly in conditions of sustained synaptic activity., Graphical Abstract, Highlights • Anchoring of KARs at glutamatergic synapses depends on receptor-glutamate binding • KARs activation/desensitization promotes receptors trapping at glutamatergic synapses • N-cadherins mediate the KAR activation/desensitization-dependent anchoring at synapses • Synaptic trapping of desensitized KARs affects short-term synaptic plasticity, Polenghi et al. report that following their activation, kainate receptors (KARs) are readily trapped at glutamatergic synapses in the open/desensitized state. This mechanism defines the number of ready-to-be-activated KARs at synapses, with direct impacts on short-term synaptic plasticity.
- Published
- 2020
- Full Text
- View/download PDF
36. Photoswitchable dynasore analogs to control endocytosis with light
- Author
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Ana Trapero, Pau Gorostiza, Alexandre Gomila-Juaneda, Artur Llobet, Ernest Giralt, Andrés Martín-Quirós, Amadeu Llebaria, Núria Camarero, Ariadna Pérez-Jiménez, Eric Macia, Jordi Hernando, and Laura Nevola
- Subjects
Transient absorption spectroscopies ,Espectroscòpia molecular ,Cell ,Spatiotemporal control ,Pharmacological properties ,010402 general chemistry ,Endocytosis ,01 natural sciences ,Flow cytometry ,03 medical and health sciences ,Photochromism ,Cellular dynamics ,Photo-switchable ,medicine ,030304 developmental biology ,Dynamin ,0303 health sciences ,Interacció cel·lular ,medicine.diagnostic_test ,Single wavelength ,Chemistry ,Micromolar concentration ,Biological activity ,General Chemistry ,Fluorescence ,Molecular spectroscopy ,0104 chemical sciences ,medicine.anatomical_structure ,Cell interaction ,Biophysics ,Small molecule inhibitor - Abstract
Altres ajuts: CERCA Programme/Generalitat de Catalunya Aquest article té una correcció a 10.1039/d0sc90189j The spatiotemporal control of cellular dynamic processes has great fundamental interest but lacks versatile molecular tools. Dynamin is a key protein in endocytosis and an appealing target to manipulate cell trafficking using patterns of light. We have developed the first photoswitchable small-molecule inhibitors of endocytosis (dynazos), by a stepwise design of the photochromic and pharmacological properties of dynasore, a dynamin inhibitor. We have characterized their photochromism with UV-visible and transient absorption spectroscopy and their biological activity using fluorescence microscopies and flow cytometry. Dynazos are water-soluble, cell permeable, and photostable, and enable fast, single-wavelength photoswitchable inhibition of clathrin-mediated endocytosis at micromolar concentration.
- Published
- 2020
37. Correction : Photoswitchable dynasore analogs to control endocytosis with light
- Author
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Ana Trapero, Ernest Giralt, Núria Camarero, Pau Gorostiza, Laura Nevola, Ariadna Pérez-Jiménez, Amadeu Llebaria, Jordi Hernando, Artur Llobet, Eric Macia, Alexandre Gomila-Juaneda, and Andrés Martín-Quirós
- Subjects
Chemistry ,Biophysics ,General Chemistry ,Endocytosis - Abstract
Correction for ‘Photoswitchable dynasore analogs to control endocytosis with light’ by Núria Camarero et al., Chem. Sci., 2020, 11, 8981–8988, DOI: 10.1039/d0sc03820b.
- Published
- 2020
38. Nanoengineered Light-Harvested Proteins for Optogenetics and Photopharmacology
- Author
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Aida Garrido-Charles, Gisela Cabré, Marta Gascón-Moya, Félix Busqué, Ramon Alibés, Jordi Hernando, and Pau Gorostiza
- Abstract
Chemical modification with nanometer precision can be used to probe and to improve the function of complex molecular entities, from organic materials to proteins and their assemblies. Using the pigment arrangement in photosynthetic light-harvesting as inspiration, we show that molecular photosensitizers can be located at well-defined distances from photoisomerizable units in proteins in order to enhance and spectrally shift their photoresponses. The approach is demonstrated in Channelrhodopsin-2 (ChR2) and in the light-gated ionotropic glutamate receptor (LiGluR), two archetypical actuators in optogenetics and photopharmacology that have been used both for fundamental and therapeutic purposes. These proof-of-concept experiments together with theoretical simulations predict that the photosensitivity can be increased several orders of magnitude using these means, thus providing a unique methodology to boost the performance of current optogenetic and photopharmacological toolboxes.
- Published
- 2019
39. Distance and Potential Dependence of Charge Transport Through the Reaction Center of Individual Photosynthetic Complexes
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Manuel López‐Ortiz, Ricardo A. Zamora, Marina Inés Giannotti, Chen Hu, Roberta Croce, Pau Gorostiza, Biophysics Photosynthesis/Energy, and LaserLaB - Energy
- Subjects
Chlorophyll ,Photosystem I Protein Complex ,photosystem I ,Proteins ,General Chemistry ,Espectroscòpia ,Spectrum analysis ,Electron Transport ,Biomaterials ,Kinetics ,current distance decay spectroscopy ,Fotosíntesi ,General Materials Science ,single molecules ,Photosynthesis ,protein electron transfer ,electrochemical scanning tunneling microscopy (ECSTM) ,Proteïnes ,Oxidation-Reduction ,Biotechnology - Abstract
Charge separation and transport through the reaction center of photosystem I (PSI) is an essential part of the photosynthetic electron transport chain. A strategy is developed to immobilize and orient PSI complexes on gold electrodes allowing to probe the complex's electron acceptor side, the chlorophyll special pair P700. Electrochemical scanning tunneling microscopy (ECSTM) imaging and current-distance spectroscopy of single protein complex shows lateral size in agreement with its known dimensions, and a PSI apparent height that depends on the probe potential revealing a gating effect in protein conductance. In current-distance spectroscopy, it is observed that the distance-decay constant of the current between PSI and the ECSTM probe depends on the sample and probe electrode potentials. The longest charge exchange distance (lowest distance-decay constant ?) is observed at sample potential 0 mV/SSC (SSC: reference electrode silver/silver chloride) and probe potential 400 mV/SSC. These potentials correspond to hole injection into an electronic state that is available in the absence of illumination. It is proposed that a pair of tryptophan residues located at the interface between P700 and the solution and known to support the hydrophobic recognition of the PSI redox partner plastocyanin, may have an additional role as hole exchange mediator in charge transport through PSI.© 2021 Wiley-VCH GmbH.
- Published
- 2021
40. Light-induced regulation of ligand-gated channel activity
- Author
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Piotr Bregestovski, Pau Gorostiza, and Galyna Maleeva
- Subjects
0301 basic medicine ,Pharmacology ,GABAA receptor ,Glutamate receptor ,Rational design ,Biology ,Ligand-gated channel activity ,03 medical and health sciences ,030104 developmental biology ,Nicotinic agonist ,Biophysics ,medicine ,Receptor ,Neuroscience ,Acetylcholine ,medicine.drug ,Acetylcholine receptor - Abstract
The control of ligand-gated receptors with light using photochromic compounds has evolved from the first handcrafted examples to accurate, engineered receptors, whose development is supported by rational design, high-resolution protein structures, comparative pharmacology and molecular biology manipulations. Photoswitchable regulators have been designed and characterized for a large number of ligand-gated receptors in the mammalian nervous system, including nicotinic acetylcholine, glutamate and GABA receptors. They provide a well-equipped toolbox to investigate synaptic and neuronal circuits in all-optical experiments. This focused review discusses the design and properties of these photoswitches, their applications and shortcomings and future perspectives in the field. Linked articles This article is part of a themed section on Nicotinic Acetylcholine Receptors. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.11/issuetoc.
- Published
- 2017
41. Fulgazepam: A Fulgimide-Based Potentiator of GABAA Receptors
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Pau Gorostiza, Burkhard Koenig, Karin Rustler, Alexandre M. J. Gomila, Galyna Maleeva, and Piotr Bregestovski
- Subjects
Agonist ,GABAA receptor ,medicine.drug_class ,Chemistry ,Allosteric regulation ,medicine ,Neurotransmission ,Potentiator ,Pharmacophore ,Receptor ,Inhibitory postsynaptic potential ,Neuroscience - Abstract
The γ-aminobutyric acid gated chloride channel represents the major mediator of inhibitory neurotransmission in the mammalian central nervous system and its dysfunction is related to severe diseases like epilepsy and depression, which can be relieved by the application of allosteric modulators. However, the drugs’ potential side-effects limit their application for long-term treatment. Applying light as external stimulus to modify the pharmacophore’s activity, as emerged in the field of photopharmacology, provides a non-invasive tool with high spatial and temporal resolution for the modulation of protein function. Herein, we report the design, synthesis, and biological evaluation of photochromic fulgimide-based benzodiazepine derivatives as light-controllable potentiators of GABAA receptors (GABAARs). A photocontrolled potentiator of GABAARs (Fulgazepam) has been identified that does not display agonist or antagonist activity and allows manipulating zebrafish larvae swimming.
- Published
- 2019
42. Photocontrol of endogenous glycine receptors in vivo
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Elvira Mukhametova, Alexandre M. J. Gomila, Karin Rustler, Piotr Bregestovski, Pau Gorostiza, Frank Peiretti, Miquel Bosch, Carme Rovira, Burkhard König, Alba Nin-Hill, Galyna Maleeva, Antoni Bautista-Barrufet, Xavier Rovira, Daniel Wutz, Marat A. Mukhamedyarov, and Mercedes Alfonso-Prieto
- Subjects
Mechanism of action ,Chemistry ,In vivo ,Allosteric regulation ,medicine ,Excitatory postsynaptic potential ,Neurotransmission ,medicine.symptom ,Receptor ,Inhibitory postsynaptic potential ,Glycine receptor ,Neuroscience - Abstract
Glycine receptors (GlyRs) are indispensable to maintain excitatory/inhibitory balance in neuronal circuits controlling reflex and rhythmic motor behaviors. Here we have developed Glyght, the first GlyR ligand controlled with light. It is selective over other cys-loop receptors, active in vivo, and displays an allosteric mechanism of action. The photomanipulation of glycinergic neurotransmission opens new avenues to understand inhibitory circuits in intact animals, and to develop drug-based phototherapies.
- Published
- 2019
- Full Text
- View/download PDF
43. Photochromic antifolate for light-activated chemotherapy
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Concepció Soler, Michela Libergoli, Núria Camarero, Alexandre M. J. Gomila, Carlo Matera, and Pau Gorostiza
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chemistry.chemical_compound ,Chemotherapy ,Photochromism ,Chemistry ,medicine.medical_treatment ,Light activated ,Antifolate ,Cancer research ,medicine - Published
- 2019
44. Reversible silencing of endogenous receptors in intact brain tissue using 2-photon pharmacology
- Author
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Hyojung Lee, Miquel Bosch, Lídia Bardia, Gertrudis Perea, Amadeu Llebaria, Pau Gorostiza, Kira E. Poskanzer, Anna Lladó, Silvia Pittolo, Julien Colombelli, Sébastien Tosi, Xavier Gómez-Santacana, Eduardo Soriano, European Commission, Ministerio de Economía y Competitividad (España), Llebaría, Amadeu, and Llebaría, Amadeu [0000-0002-8200-4827]
- Subjects
0301 basic medicine ,Photoactivation ,Allosteric modulator ,Receptor, Metabotropic Glutamate 5 ,Photopharmacology ,Endogeny ,Receptors, Cell Surface ,Pharmacology ,Rats, Sprague-Dawley ,03 medical and health sciences ,0302 clinical medicine ,Pharmacological selectivity ,Receptors ,MD Multidisciplinary ,Gene silencing ,Animals ,Functional silencing ,Receptor ,2-photon pharmacology ,Neurons ,Photons ,Multidisciplinary ,Chemistry ,Glutamate receptor ,Brain ,Phenotype ,Metabotropic Glutamate 5 ,3. Good health ,Rats ,Optogenetics ,030104 developmental biology ,Metabotropic receptor ,PNAS Plus ,Astrocytes ,Cell Surface ,Calcium ,Sprague-Dawley ,Signal transduction ,030217 neurology & neurosurgery - Abstract
The physiological activity of proteins is often studied with loss-of-function genetic approaches, but the corresponding phenotypes develop slowly and can be confounding. Photopharmacology allows direct, fast, and reversible control of endogenous protein activity, with spatiotemporal resolution set by the illumination method. Here, we combine a photoswitchable allosteric modulator (alloswitch) and 2-photon excitation using pulsed near-infrared lasers to reversibly silence metabotropic glutamate 5 (mGlu5) receptor activity in intact brain tissue. Endogenous receptors can be photoactivated in neurons and astrocytes with pharmacological selectivity and with an axial resolution between 5 and 10 μm. Thus, 2-photon pharmacology using alloswitch allows investigating mGlu5-dependent processes in wild-type animals, including synaptic formation and plasticity, and signaling pathways from intracellular organelles. © 2019 National Academy of Sciences. All rights reserved., ACKNOWLEDGMENTS. We thank Jordi Hernando (Autonomous University of Barcelona) for useful discussions on 2-photon excitation; Pere Català (Utrecht University) for help with GCaMP; Francisco Ciruela (University of Barcelona) for mGlu5-eYFP plasmid; Erin Schuman and Stephan Junek (Max Planck Institute for Brain Research, Frankfurt) for preliminary 2-photon excitation experiments; and Ashraf Muhaisen (University of Barcelona) for help with slicing. This research received funding from European Union Research and Innovation Programme Horizon 2020 [Human Brain Project SGA2 Grant Agreement 785907 (WaveScalES)], European Research ERA-Net SynBio programme (Modulightor project), and financial support from Agency for Management of University and Research Grants/Generalitat de Catalunya (CERCA Programme; 2017-SGR-1442 project), Fonds Européen de Développement Économique et Régional (FEDER) funds, Ministry of Economy and Competitiveness (MINECO)/FEDER (Grant CTQ2016-80066-R), and the Fundaluce foundation. S.P. was supported by an FI fellowship from the Agency for Management of University and Research Grants/Generalitat de Catalunya (2014FI_B2 00160). H.L. was supported by an Institute for Bioengineering of Catalonia Severo Ochoa International PhD Programme fellowship from MINECO. M.B. was supported by a H2020-MSCA-IF Reintegration Grant. K.E.P. receives support from NIH/National Institute of Neurological Disorders and Stroke Grant R01NS099254 and NSF Biophotonics Grant 1604544. E.S. receives support from MINECO (Grant SAF2016-7426).
- Published
- 2019
45. Electrochemically Gated Long Distance Charge Transport in Photosystem I
- Author
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Manuel López Ortiz, Maria Elena Antinori, Emilie Wientjes, Pau Gorostiza, Ismael Díez-Pérez, Roberta Croce, and Montserrat López Martínez
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Electron transfer ,Materials science ,P700 ,law ,Chemical physics ,Fermi energy ,Scanning tunneling microscope ,Electrochemistry ,Photosystem I ,Electron transport chain ,Redox ,law.invention - Abstract
The transport of electrons along photosynthetic and respiratory chains involves a series of enzymatic reactions that are coupled through redox mediators, including proteins and small molecules. The use of native and synthetic redox probes is key to understand charge transport mechanisms, and to design bioelectronic sensors and solar energy conversion devices. However, redox probes have limited tunability to exchange charge at the desired electrochemical potentials (energy levels) and at different protein sites. Here, we take advantage of electrochemical scanning tunneling microscopy (ECSTM) to control the Fermi energy and nanometric position of the ECSTM probe in order to study electron transport in individual photosystem I (PSI) complexes. Current-distance measurements at different potentiostatic conditions indicate that PSI supports long-distance transport that is electrochemically gated near the redox potential of P700, with current extending farther under hole injection conditions.
- Published
- 2019
46. Control of Cardiac Function in vivo with a Light-Regulated Drug
- Author
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Carlo Matera, Alexandre M. J. Gomila, Ulrike Holzgrabe, Enrique Claro, Eduard Guasch, Fabio Riefolo, Pau Gorostiza, Michael Decker, Aida Garrido-Charles, Montserrat Batlle, Roser Masgrau, and Luca Agnetta
- Subjects
Agonist ,Calcium imaging ,In vivo ,Chemistry ,medicine.drug_class ,Allosteric regulation ,Muscarinic acetylcholine receptor ,medicine ,Cholinergic ,Optogenetics ,Receptor ,Cell biology ,3. Good health - Abstract
Remote control of physiological functions with light offers the promise of unveiling their complex spatiotemporal dynamics in vivo, and enabling highly focalized therapeutic interventions with reduced systemic toxicity. Optogenetic methods have been implemented in the heart, but the need of genetic manipulation jeopardizes clinical applicability. This study aims at developing, testing and validating the first light-regulated drug with cardiac effects, in order to avoid the requirement of genetic manipulation offered by optogenetic methods. A M2 muscarinic acetylcholine receptors (mAChRs) light-regulated drug (PAI) was designed, synthesized and pharmacologically characterized. The design was based on the orthosteric mAChRs agonist Iperoxo, an allosteric M2 ligand, and a photoswitchable azobenzene linker. PAI can be reversibly photoisomerized between cis and trans configurations under ultraviolet (UV) and visible light, respectively, and it reversibly photoswitches the activity of M2 muscarinic acetylcholine receptors. We have evaluated in vitro photoresponses using a calcium imaging assay in genetically unmodified receptors overexpressed in mammalian cells. Furthermore, using this new chemical tool, we demonstrate for the first time photoregulation of cardiac function in vivo in wildtype frog tadpoles and in rats with a method that does not require genetic manipulation. Such a new approach may enable enhanced spatial and temporal selectivity for cardiovascular drugs.
- Published
- 2018
47. A Photoswitchable Antimetabolite for Targeted Photoactivated Chemotherapy
- Author
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Alexandre M. J. Gomila, Michela Libergoli, Pau Gorostiza, Concepció Soler, Carlo Matera, Núria Camarero, and Universitat de Barcelona
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Models, Molecular ,0301 basic medicine ,medicine.medical_treatment ,Photodynamic therapy ,Pharmacology ,01 natural sciences ,Biochemistry ,chemistry.chemical_compound ,Colloid and Surface Chemistry ,Dihydrofolate reductase ,Enzyme Inhibitors ,Càncer ,Zebrafish ,Cancer ,Molecular Structure ,biology ,Chemistry ,Fototeràpia ,Photochemical Processes ,3. Good health ,Nanomedicine ,Tolerability ,Antifolate ,Nanomedicina ,medicine.drug ,medicine.drug_class ,Antineoplastic Agents ,Artritis reumatoide ,Antimetabolite ,Catalysis ,Structure-Activity Relationship ,03 medical and health sciences ,Fotoquimioteràpia ,In vivo ,medicine ,Animals ,Humans ,Psoriasis ,Rheumatoid arthritis ,Cell Proliferation ,Psoriasi ,Chemotherapy ,Dose-Response Relationship, Drug ,010405 organic chemistry ,General Chemistry ,Phototherapy ,0104 chemical sciences ,Tetrahydrofolate Dehydrogenase ,Methotrexate ,030104 developmental biology ,Photochemotherapy ,biology.protein ,Drug Screening Assays, Antitumor - Abstract
The efficacy and tolerability of systemically administered anticancer agents are limited by their off-target effects. Precise spatiotemporal control over their cytotoxic activity would allow improving chemotherapy treatments, and light-regulated drugs are well suited to this purpose. We have developed phototrexate, the first photoswitchable inhibitor of the human dihydrofolate reductase (DHFR), as a photochromic analogue of methotrexate, a widely prescribed chemotherapeutic drug to treat cancer and psoriasis. Quantification of the light-regulated DHFR enzymatic activity, cell proliferation, and in vivo effects in zebrafish show that phototrexate behaves as a potent antifolate in its photoactivated cis configuration and that it is nearly inactive in its dark-relaxed trans form. Thus, phototrexate constitutes a proof-of-concept to design light-regulated cytotoxic small molecules and a step forward to develop targeted anticancer photochemotherapies with localized efficacy and reduced adverse effects.
- Published
- 2018
48. OptoGluNAM4.1, a Photoswitchable Allosteric Antagonist for Real-Time Control of mGlu 4 Receptor Activity
- Author
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Amadeu Llebaria, Jesús Giraldo, Pau Gorostiza, Xavier Rovira, Charleine Zussy, Ana Trapero, Silvia Pittolo, Chris Jopling, Adèle Faucherre, Cyril Goudet, Jean-Philippe Pin, European Research Council, Trapero, Ana [0000-0003-4526-7895], Llebaría, Amadeu [0000-0002-8200-4827], Institut de Génomique Fonctionnelle (IGF), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), Institute for Bioengineering of Catalonia [Barcelona] (IBEC), Trapero, Ana, and Llebaría, Amadeu
- Subjects
0301 basic medicine ,Time Factors ,Light ,[SDV]Life Sciences [q-bio] ,Clinical Biochemistry ,Chronic pain ,Pharmacology ,Receptors, Metabotropic Glutamate ,Biochemistry ,Mice ,0302 clinical medicine ,Drug Discovery ,ComputingMilieux_MISCELLANEOUS ,Zebrafish ,Azobenzene ,Metabotropic glutamate receptor 5 ,Metabotropic glutamate receptor 4 ,mGlu receptor ,Metabotropic glutamate receptor 7 ,Metabotropic glutamate receptor 6 ,Dolor crònic ,Pyrrolidonecarboxylic Acid ,3. Good health ,Cell biology ,Molecular Medicine ,Metabotropic glutamate receptor 1 ,Metabotropic glutamate receptor 2 ,Nervous system ,Pain Threshold ,Allosteric modulator ,OptoGluNAM4.1 ,Biology ,Structure-Activity Relationship ,03 medical and health sciences ,Allosteric Regulation ,Enzyme-linked receptor ,Animals ,Humans ,Sistema nerviós ,Molecular Biology ,Dose-Response Relationship, Drug ,Mice, Inbred C57BL ,Disease Models, Animal ,HEK293 Cells ,030104 developmental biology ,Schizophrenia ,Azo Compounds ,030217 neurology & neurosurgery - Abstract
OptoGluNAM4.1, a negative allosteric modulator (NAM) of metabotropic glutamate receptor 4 (mGlu4) contains a reactive group that covalently binds to the receptor and a blue-light-activated, fast-relaxing azobenzene group that allows reversible receptor activity photocontrol in vitro and in vivo. OptoGluNAM4.1 induces light-dependent behavior in zebrafish and reverses the activity of the mGlu4 agonist LSP4-2022 in a mice model of chronic pain, defining a photopharmacological tool to better elucidate the physiological roles of the mGlu4 receptor in the nervous system. © 2016 Elsevier Ltd, We are grateful to C. Serra, L. Muñoz, J. Hernando, F. Malhaire, Y. Pérez, M. Izquierdo-Serra, F. Aguado, S. Laffray, E. Bourinet, F.Codony (GenIUL), and Y. Chomis (Viewpoint) for helpful discussions and technical support. A.F. is supported by a Fondation Lefoulon Delalande postdoctoral fellowship , C.J. by a grant from INSERM ATIP-AVENIR and Marie Curie CIG ( PCIG12-GA-2012-332772 ). A.F. and C.J. are members of the Laboratory of Excellence Ion Channel Science and Therapeutics supported by a grant from the ANR . J.-P.P. is a member of the Laboratory of Excellence Epingenmed. We acknowledge financial support from the European Union's Seventh Framework Program for research, technological development, and demonstration under grant agreements 270483 (Focus), 210355 (Opticalbullet), and 335011 (Theralight) to P.G.; the Federation of European Biochemical Societies ; the Catalan government ( 2012FI_B 01122 to S.P., 2014SGR-1251 to P.G., and 2014SGR-0109 to A. Llebaria); the Spanish Government ( SAF2014-58396-R to J.G., CTQ2014-57020-R to A.L., and CTQ2013-43892R to P.G.); the ERANET Neuron LIGHTPAIN project (to A.L., J.G., and J.-P.P.); the Ramón Areces Foundation , the ERANET SynBio Modulightor project (to P.G.); the HBP Wavescales project (to P.G.); the Fondation Recherche Médicale (FRM team DEQ20130326522 to J.P.P.); the Agence Nationale de la Recherche ( ANR-13-BSV1-006 to C.G.) and the Beatriu de Pinós program of Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR) for the support of X.R.
- Published
- 2016
49. Dendrimer-based Uneven Nanopatterns to Locally Control Surface Adhesiveness: A Method to Direct Chondrogenic Differentiation
- Author
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Ezequiel Perez-Inestrosa, Ignasi Casanellas, José A. Andrades, Daniel Collado, Iro Tsintzou, Cristina Rodríguez-Pereira, Yolanda Vida, José Becerra, Josep Samitier, Pau Gorostiza, Anna Lagunas, Joana Magalhães, [Casanellas, Ignasi] BIST, Inst Bioengn Catalonia IBEC, Barcelona, Spain, [Lagunas, Anna] BIST, Inst Bioengn Catalonia IBEC, Barcelona, Spain, [Tsintzou, Iro] BIST, Inst Bioengn Catalonia IBEC, Barcelona, Spain, [Samitier, Josep] BIST, Inst Bioengn Catalonia IBEC, Barcelona, Spain, [Casanellas, Ignasi] Univ Barcelona, Dept Engn Elect, Barcelona, Spain, [Samitier, Josep] Univ Barcelona, Dept Engn Elect, Barcelona, Spain, [Lagunas, Anna] Networking Biomed Res Ctr CIBER, Madrid, Spain, [Magalhaes, Joana] Networking Biomed Res Ctr CIBER, Madrid, Spain, [Gorostiza, Pau] Networking Biomed Res Ctr CIBER, Madrid, Spain, [Andrades, Jose A.] Networking Biomed Res Ctr CIBER, Madrid, Spain, [Becerra, Jose] Networking Biomed Res Ctr CIBER, Madrid, Spain, [Samitier, Josep] Networking Biomed Res Ctr CIBER, Madrid, Spain, [Vida, Yolanda] Univ Malaga UMA, Dept Organ Chem, Inst Invest Biomed Malaga IBIMA, Malaga, Spain, [Collado, Daniel] Univ Malaga UMA, Dept Organ Chem, Inst Invest Biomed Malaga IBIMA, Malaga, Spain, [Perez-Inestrosa, Ezequiel] Univ Malaga UMA, Dept Organ Chem, Inst Invest Biomed Malaga IBIMA, Malaga, Spain, [Vida, Yolanda] Andalusian Ctr Nanomed & Biotechnol BIONAND, Malaga, Spain, [Collado, Daniel] Andalusian Ctr Nanomed & Biotechnol BIONAND, Malaga, Spain, [Perez-Inestrosa, Ezequiel] Andalusian Ctr Nanomed & Biotechnol BIONAND, Malaga, Spain, [Becerra, Jose] Andalusian Ctr Nanomed & Biotechnol BIONAND, Malaga, Spain, [Rodriguez-Pereira, Cristina] UDC, Sergas,CHUAC, Grp Reumatol,Inst Invest Biomed A Coruna INIBIC, Unidad Bioingn Tisular & Terapia Celular GBTTC CH, La Coruna, Spain, [Magalhaes, Joana] UDC, Sergas,CHUAC, Grp Reumatol,Inst Invest Biomed A Coruna INIBIC, Unidad Bioingn Tisular & Terapia Celular GBTTC CH, La Coruna, Spain, [Gorostiza, Pau] ICREA, Barcelona, Spain, [Andrades, Jose A.] Univ Malaga UMA, Dept Cell Biol Genet & Physiol, Inst Invest Biomed Malaga IBIMA, Malaga, Spain, [Becerra, Jose] Univ Malaga UMA, Dept Cell Biol Genet & Physiol, Inst Invest Biomed Malaga IBIMA, Malaga, Spain, Networking Biomedical Research Center (CIBER), Spain, VI National RDi Plan, Iniciativa Ingenio, Consolider Program, CIBER Actions, Instituto de Salud Carlos III, European Regional Development Fund, Commission for Universities and Research of the Department of Innovation, Universities, Enterprise of the Generalitat de Catalunya, projects OLIGOCODES, Spanish Ministry of Economy and Competitiveness, and INTERREG V-A Spain-Portugal POCTEP
- Subjects
General Chemical Engineering ,Nanopattern ,Density ,Growth ,02 engineering and technology ,Cell-adhesion ,01 natural sciences ,law.invention ,Contact angle ,Mice ,law ,Arginine-Glycine-Aspartic Acid (RGD) ,Dendrimer ,General Neuroscience ,Adhesiveness ,021001 nanoscience & nanotechnology ,Stem-cells ,Issue 131 ,Collagen ,Scanning tunneling microscope ,0210 nano-technology ,Chondrogenesis ,Oligopeptides ,D-glucose ,Dendrimers ,Materials science ,Surface Properties ,Atomic Force Microscopy (AFM) ,Bioengineering ,010402 general chemistry ,General Biochemistry, Genetics and Molecular Biology ,Immobilization ,Scanning probe microscopy ,Adsorption ,X-ray photoelectron spectroscopy ,Cell Adhesion ,Animals ,Cell adhesion ,Fibronectin ,Nanoscopic scale ,General Immunology and Microbiology ,Mesenchymal Stem Cells (Mscs) ,Mesenchymal Stem Cells ,Fibroblasts ,Nanostructures ,0104 chemical sciences ,NIH 3T3 Cells ,Biophysics - Abstract
Cellular adhesion and differentiation is conditioned by the nanoscale disposition of the extracellular matrix (ECM) components, with local concentrations having a major effect. Here we present a method to obtain large-scale uneven nanopatterns of arginine-glycine-aspartic acid (RGD)-functionalized dendrimers that permit the nanoscale control of local RGD surface density. Nanopatterns are formed by surface adsorption of dendrimers from solutions at different initial concentrations and are characterized by water contact angle (CA), X-ray photoelectron spectroscopy (XPS), and scanning probe microscopy techniques such as scanning tunneling microscopy (STM) and atomic force microscopy (AFM). The local surface density of RGD is measured using AFM images by means of probability contour maps of minimum interparticle distances and then correlated with cell adhesion response and differentiation. The nanopatterning method presented here is a simple procedure that can be scaled up in a straightforward manner to large surface areas. It is thus fully compatible with cell culture protocols and can be applied to other ligands that exert concentration-dependent effects on cells.
- Published
- 2018
50. Long distance electron transfer through the aqueous solution between redox partner proteins
- Author
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Miguel A. De la Rosa, Alba Nin-Hill, Irene Díaz-Moreno, Pau Gorostiza, Anna Lagunas, Alejandra Guerra-Castellano, Josep Samitier, Carme Rovira, Universidad de Sevilla. Departamento de Bioquímica Vegetal y Biología Molecular, Ministerio de Ciencia, Innovación y Universidades (España), Ministerio de Economía y Competitividad (España), Junta de Andalucía, Generalitat de Catalunya, Centros de Investigación Biomédica en Red (España), European Commission, and Instituto de Salud Carlos III
- Subjects
0301 basic medicine ,Science ,Cell Respiration ,Arabidopsis ,General Physics and Astronomy ,Ionic bonding ,Electrons ,Molecular Dynamics Simulation ,Electrochemistry ,Redox ,Article ,Biophysical Phenomena ,General Biochemistry, Genetics and Molecular Biology ,Electron Transport ,03 medical and health sciences ,Electron transfer ,Molecular dynamics ,Escherichia coli ,Photosynthesis ,lcsh:Science ,Electrochemical potential ,Multidisciplinary ,Aqueous solution ,Chemistry ,Spectrum Analysis ,Proteins ,General Chemistry ,Electron transport chain ,030104 developmental biology ,Chemical physics ,lcsh:Q ,Oxidation-Reduction - Abstract
Despite the importance of electron transfer (ET) between redox proteins in photosynthesis and respiration, the inter-protein ET rate between redox partner proteins has never been measured as a function of their separation in aqueous solution. Here we use electrochemical tunneling spectroscopy to show that the current between two protein partners decays along more than 10 nm in the solution. Molecular dynamics simulations reveal a reduced ionic density and extended electric field in the volume confined between the proteins. The distance-decay factor and the calculated local barrier for ET are regulated by the electrochemical potential applied to the proteins. Redox partners could use electrochemically gated, long distance ET through the solution in order to conciliate high ET specificity with weak binding, thus keeping high turnover rates in the crowded environment of cells., The project was funded by CERCA, FEDER, and Networking Biomedical Research Center (CIBER), Spain. CIBER is an initiative funded by the VI National R&D&i Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions, and the Instituto de Salud Carlos III, with the support of the European Regional Development Fund. This work was financially supported by the Commission for Universities and Research of the Department of Innovation, Universities, and Enterprise of the Generalitat de Catalunya (2017 SGR 1442, 2014 SGR 1442, and 2017 SGR 1189), by the Andalusian Government (BIO198), and the projects CTQ2016–80066-R, CTQ2015-66194-R, CTQ2017-85496-P and Spanish Structures of Excellence Maria de Maeztu, MDM2017-0767, and BFU2015-71017-P/BMC awarded by the Spanish Ministry of Economy and Competitiveness. A.N.-H. thanks the Generalitat de Catalunya-AGAUR for a FI scholarship. MINECO/ICTI2013-2016/CTQ2015-66194-R MINECO/ICTI2013-2016/BFU2015-71017-P MICIU/ICTI2017-2020/CTQ2017-85496-P MICIU/ICTI2017-2020/MDM2017-0767
- Published
- 2018
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