36 results on '"Polyhach Y"'
Search Results
2. The 3D solution structure of discoidal high-density lipoprotein particles
- Author
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Bibow, S., primary, Polyhach, Y., additional, Eichmann, C., additional, Chi, C.N., additional, Kowal, J., additional, Stahlberg, H., additional, Jeschke, G., additional, Guentert, P., additional, and Riek, R., additional
- Published
- 2016
- Full Text
- View/download PDF
3. Distance determination between low-spin ferric haem and nitroxide spin label using DEER: the neuroglobin case
- Author
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Ezhevskaya, M., primary, Bordignon, E., additional, Polyhach, Y., additional, Moens, L., additional, Dewilde, S., additional, Jeschke, G., additional, and Van Doorslaer, S., additional
- Published
- 2013
- Full Text
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4. Structure and conformational dynamics of the sodium/proline transporter PutP based on protein chemical and EPR spectroscopic analyses
- Author
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Hilger, D., primary, Raba, M., additional, Lipiszko, K., additional, Polyhach, Y., additional, Jeschke, G., additional, Dunkel, S., additional, Klare, J., additional, Steinhoff, H.-J., additional, Bracher, S., additional, Quick, Matthias, additional, and Jung, H., additional
- Published
- 2012
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5. High-Resolution Structure of a Na+/H+ Antiporter Dimer Obtained by Pulsed Electron Paramagnetic Resonance Distance Measurements
- Author
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Hilger, D., primary, Polyhach, Y., additional, Padan, E., additional, Jung, H., additional, and Jeschke, G., additional
- Published
- 2007
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6. Native defects and rare-earth impurities interaction in IV-VI crystals
- Author
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Zayachuk, D., Polyhach, Y., Slynko, E., Khandozhko, O., Kempnyk, V., and Baltrunas, D.
- Published
- 2001
- Full Text
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7. Intermolecular contributions, filtration effects and signal composition of SIFTER (single-frequency technique for refocusing).
- Author
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Vanas A, Soetbeer J, Breitgoff FD, Hintz H, Sajid M, Polyhach Y, Godt A, Jeschke G, Yulikov M, and Klose D
- Abstract
To characterize structure and molecular order in the nanometre range, distances between electron spins and their distributions can be measured via dipolar spin-spin interactions by different pulsed electron paramagnetic resonance experiments. Here, for the single-frequency technique for refocusing dipolar couplings (SIFTER), the buildup of dipolar modulation signal and intermolecular contributions is analysed for a uniform random distribution of monoradicals and biradicals in frozen glassy solvent by using the product operator formalism for electron spin S = 1 / 2 . A dipolar oscillation artefact appearing at both ends of the SIFTER time trace is predicted, which originates from the weak coherence transfer between biradicals. The relative intensity of this artefact is predicted to be temperature independent but to increase with the spin concentration in the sample. Different compositions of the intermolecular background are predicted in the case of biradicals and in the case of monoradicals. Our theoretical account suggests that the appropriate procedure of extracting the intramolecular dipolar contribution (form factor) requires fitting and subtracting the unmodulated part, followed by division by an intermolecular background function that is different in shape. This scheme differs from the previously used heuristic background division approach. We compare our theoretical derivations to experimental SIFTER traces for nitroxide and trityl monoradicals and biradicals. Our analysis demonstrates a good qualitative match with the proposed theoretical description. The resulting perspectives for a quantitative analysis of SIFTER data are discussed., Competing Interests: The contact author has declared that none of the authors has any competing interests., (Copyright: © 2023 Agathe Vanas et al.)
- Published
- 2023
- Full Text
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8. Ligand-specific conformational change drives interdomain allostery in Pin1.
- Author
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Born A, Soetbeer J, Henen MA, Breitgoff F, Polyhach Y, Jeschke G, and Vögeli B
- Subjects
- Allosteric Regulation, Binding Sites, Catalytic Domain, Ligands, Protein Binding, Protein Domains, NIMA-Interacting Peptidylprolyl Isomerase metabolism
- Abstract
Pin1 is a two-domain cell regulator that isomerizes peptidyl-prolines. The catalytic domain (PPIase) and the other ligand-binding domain (WW) sample extended and compact conformations. Ligand binding changes the equilibrium of the interdomain conformations, but the conformational changes that lead to the altered domain sampling were unknown. Prior evidence has supported an interdomain allosteric mechanism. We recently introduced a magnetic resonance-based protocol that allowed us to determine the coupling of intra- and interdomain structural sampling in apo Pin1. Here, we describe ligand-specific conformational changes that occur upon binding of pCDC25c and FFpSPR. pCDC25c binding doubles the population of the extended states compared to the virtually identical populations of the apo and FFpSPR-bound forms. pCDC25c binding to the WW domain triggers conformational changes to propagate via the interdomain interface to the catalytic site, while FFpSPR binding displaces a helix in the PPIase that leads to repositioning of the PPIase catalytic loop., (© 2022. The Author(s).)
- Published
- 2022
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9. Regularized dynamical decoupling noise spectroscopy - a decoherence descriptor for radicals in glassy matrices.
- Author
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Soetbeer J, Ibáñez LF, Berkson Z, Polyhach Y, and Jeschke G
- Abstract
Decoherence arises from a fluctuating spin environment, captured by its noise spectrum S ( ω ). Dynamical decoupling (DD) with n π pulses extends the dephasing time if the associated filter function attenuates S ( ω ). Inversely, DD noise spectroscopy (DDNS) reconstructs S ( ω ) from DD data by approximating the filters pass band by a δ -function. This restricts application to qubit-like spin systems with inherently long dephasing times and/or many applicable pulses. We introduce regularized DDNS to lift this limitation and thereby infer S ( ω ) from DD traces of paramagnetic centers in glassy o -terphenyl and water-glycerol matrices recorded with n ≤ 5. For nitroxide radicals at low temperatures, we utilize deuteration to identify distinct matrix- and spin center-induced spectral features. The former extends up to a matrix-specific cut-off frequency and characterizes nuclear spin diffusion. We demonstrate that rotational tunneling of intramolecular methyl groups drives the latter process, whereas at elevated temperatures S ( ω ) reflects the classical methyl group reorientation. Ultimately, S ( ω ) visualizes and quantifies variations in the electron spins couplings and thus reports on the underlying spin dynamics as a powerful decoherence descriptor.
- Published
- 2021
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10. Reconstruction of Coupled Intra- and Interdomain Protein Motion from Nuclear and Electron Magnetic Resonance.
- Author
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Born A, Soetbeer J, Breitgoff F, Henen MA, Sgourakis N, Polyhach Y, Nichols PJ, Strotz D, Jeschke G, and Vögeli B
- Subjects
- Humans, Models, Molecular, Molecular Dynamics Simulation, Nitric Oxide Synthase Type III chemistry, Nitric Oxide Synthase Type III metabolism, Protein Conformation, Magnetic Resonance Spectroscopy methods, NIMA-Interacting Peptidylprolyl Isomerase chemistry
- Abstract
Proteins composed of multiple domains allow for structural heterogeneity and interdomain dynamics that may be vital for function. Intradomain structures and dynamics can influence interdomain conformations and vice versa . However, no established structure determination method is currently available that can probe the coupling of these motions. The protein Pin1 contains separate regulatory and catalytic domains that sample "extended" and "compact" states, and ligand binding changes this equilibrium. Ligand binding and interdomain distance have been shown to impact the activity of Pin1, suggesting interdomain allostery. In order to characterize the conformational equilibrium of Pin1, we describe a novel method to model the coupling between intra- and interdomain dynamics at atomic resolution using multistate ensembles. The method uses time-averaged nuclear magnetic resonance (NMR) restraints and double electron-electron resonance (DEER) data that resolve distance distributions. While the intradomain calculation is primarily driven by exact nuclear Overhauser enhancements (eNOEs), J couplings, and residual dipolar couplings (RDCs), the relative domain distribution is driven by paramagnetic relaxation enhancement (PREs), RDCs, interdomain NOEs, and DEER. Our data support a 70:30 population of the compact and extended states in apo Pin1. A multistate ensemble describes these conformations simultaneously, with distinct conformational differences located in the interdomain interface stabilizing the compact or extended states. We also describe correlated conformations between the catalytic site and interdomain interface that may explain allostery driven by interdomain contact.
- Published
- 2021
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11. Gradual opening of Smc arms in prokaryotic condensin.
- Author
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Vazquez Nunez R, Polyhach Y, Soh YM, Jeschke G, and Gruber S
- Subjects
- Humans, Adenosine Triphosphatases metabolism, Bacterial Proteins metabolism, Cell Cycle Proteins metabolism, DNA-Binding Proteins metabolism, Multiprotein Complexes metabolism, Prokaryotic Cells metabolism
- Abstract
Multi-subunit SMC ATPases control chromosome superstructure apparently by catalyzing a DNA-loop-extrusion reaction. SMC proteins harbor an ABC-type ATPase "head" and a "hinge" dimerization domain connected by a coiled coil "arm." Two arms in a SMC dimer can co-align, thereby forming a rod-shaped particle. Upon ATP binding, SMC heads engage, and arms are thought to separate. Here, we study the shape of Bacillus subtilis Smc-ScpAB by electron-spin resonance spectroscopy. Arm separation is readily detected proximal to the heads in the absence of ligands, and separation near the hinge largely depends on ATP and DNA. Artificial blockage of arm opening eliminates DNA stimulation of ATP hydrolysis but does not prevent basal ATPase activity. We report an arm contact as being important for controlling the transformations. Point mutations at this arm interface eliminated Smc function. We propose that partially open, intermediary conformations provide directionality to SMC DNA translocation by (un)binding suitable DNA substrates., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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12. Dynamical decoupling in water-glycerol glasses: a comparison of nitroxides, trityl radicals and gadolinium complexes.
- Author
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Soetbeer J, Millen M, Zouboulis K, Hülsmann M, Godt A, Polyhach Y, and Jeschke G
- Abstract
Our previous study on nitroxides in o-terphenyl (OTP) revealed two separable decoherence processes at low temperatures, best captured by the sum of two stretched exponentials (SSE) model. Dynamical decoupling (DD) extends both associated dephasing times linearly for 1 to 5 refocusing pulses [Soetbeer et al., Phys. Chem. Chem. Phys., 2018, 20, 1615]. Here we demonstrate an analogous DD behavior of water-soluble nitroxides in water-glycerol glass by using nitroxide and/or solvent deuteration for component assignment. Compared to the conventional Hahn experiment, we show that Carr-Purcell and Uhrig DD schemes are superior in resolving and identifying active dephasing mechanisms. Thereby, we observe a partial coherence loss to intramolecular nitroxide and trityl nuclei that can be alleviated, while the zero field splitting-induced losses for gadolinium labels cannot be refocused and contribute even at the central transition of this spin-7/2 system. Independent of the studied spin system, Uhrig DD leads to a characteristic convex dephasing envelope in both protonated water-glycerol and OTP glass, thus outperforming the Carr-Purcell scheme.
- Published
- 2021
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13. Quantification of Redox Sites during Catalytic Propane Oxychlorination by Operando EPR Spectroscopy.
- Author
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Zichittella G, Polyhach Y, Tschaggelar R, Jeschke G, and Pérez-Ramírez J
- Abstract
Identification and quantification of redox-active centers at relevant conditions for catalysis is pivotal to understand reaction mechanisms and requires development of advanced operando methods. Herein, we demonstrate operando EPR spectroscopy as an important technique to quantify the oxidation state of representative CrPO
4 and EuOCl catalysts during propane oxychlorination, an attractive route for propylene production. In particular, we show that the space-time-yield of C3 H6 correlates with the amount of Cr2+ and Eu2+ ions generated over the catalysts during reaction. These results provide a powerful strategy to gather quantitative understanding of selective alkane oxidation, which could potentially be extrapolated to other functionalization approaches and operating conditions., (© 2020 Wiley-VCH GmbH.)- Published
- 2021
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14. Linear and Kinked Oligo(phenyleneethynylene)s as Ideal Molecular Calibrants for Förster Resonance Energy Transfer.
- Author
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Czar MF, Breitgoff FD, Sahoo D, Sajid M, Ramezanian N, Polyhach Y, Jeschke G, Godt A, and Zenobi R
- Abstract
We show that oligo(phenyleneethynylene)s (oligoPEs) are ideal spacers for calibrating dye pairs used for Förster resonance energy transfer (FRET). Ensemble FRET measurements on linear and kinked diads with such spacers show the expected distance and orientation dependence of FRET. Measured FRET efficiencies match excellently with those predicted using a harmonic segmented chain model, which was validated by end-to-end distance distributions obtained from pulsed electron paramagnetic resonance measurements on spin-labeled oligoPEs with comparable label distances.
- Published
- 2019
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15. Comparison of the functional properties of trimeric and monomeric CaiT of Escherichia coli.
- Author
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Bracher S, Hilger D, Guérin K, Polyhach Y, Jeschke G, Krafczyk R, Giacomelli G, and Jung H
- Subjects
- Amino Acid Substitution, Antiporters genetics, Cell Membrane metabolism, Chromatography, Gel, Cysteine genetics, Detergents chemistry, Escherichia coli genetics, Escherichia coli metabolism, Escherichia coli Proteins genetics, Protein Multimerization, Tryptophan chemistry, Antiporters chemistry, Antiporters metabolism, Escherichia coli Proteins chemistry, Escherichia coli Proteins metabolism
- Abstract
Secondary transporters exist as monomers, dimers or higher state oligomers. The significance of the oligomeric state is only partially understood. Here, the significance of the trimeric state of the L-carnitine/γ-butyrobetaine antiporter CaiT of Escherichia coli was investigated. Amino acids important for trimer stability were identified and experimentally verified. Among others, CaiT-D288A and -D288R proved to be mostly monomeric in detergent solution and after reconstitution into proteoliposomes, as shown by blue native gel electrophoresis, gel filtration, and determination of intermolecular distances. CaiT-D288A was fully functional with kinetic parameters similar to the trimeric wild-type. Significant differences in amount and stability in the cell membrane between monomeric and trimeric CaiT were not observed. Contrary to trimeric CaiT, addition of substrate had no or only a minor effect on the tryptophan fluorescence of monomeric CaiT. The results suggest that physical contacts between protomers are important for the substrate-induced changes in protein fluorescence and the underlying conformational alterations.
- Published
- 2019
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16. EPR Techniques to Probe Insertion and Conformation of Spin-Labeled Proteins in Lipid Bilayers.
- Author
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Bordignon E, Kucher S, and Polyhach Y
- Subjects
- Gadolinium chemistry, Liposomes chemistry, Magnetic Resonance Spectroscopy methods, Molecular Conformation, Mutagenesis, Site-Directed methods, Spin Labels, Temperature, Water chemistry, Electron Spin Resonance Spectroscopy methods, Lipid Bilayers chemistry, Membrane Proteins chemistry
- Abstract
Electron paramagnetic resonance (EPR) spectroscopy of spin-labeled membrane proteins is a valuable biophysical technique to study structural details and conformational transitions of proteins close to their physiological environment, for example, in liposomes, membrane bilayers, and nanodiscs. Unlike in nuclear magnetic resonance (NMR) spectroscopy, having only one or few specific side chains labeled at a time with paramagnetic probes makes the size of the object under investigation irrelevant in terms of technique sensitivity. As a drawback, extensive site-directed mutagenesis is required in order to analyze the properties of the protein under investigation. EPR can provide detailed information on side chain dynamics of large membrane proteins or protein complexes embedded in membranes with an exquisite sensitivity for flexible regions and on water accessibility profiles across the membrane bilayer. Moreover, distances between the two spin-labeled side chains in membrane proteins can be detected with high precision at cryogenic temperatures. The application of EPR to membrane proteins still presents some challenges in terms of sample preparation, sensitivity and data interpretation, thus it is difficult to give ready-to-go methodological recipes. However, new technological developments (arbitrary waveform generators) and new spin labels spectroscopically orthogonal to nitroxides increased the range of applicability from in vitro toward in-cell EPR experiments. This chapter is an updated version of the one published in the first edition of the book and describes the state of the art in the application of nitroxide-based site-directed spin labeling EPR to membrane proteins, addressing new tools such as arbitrary waveform generators and spectroscopically orthogonal labels, such as Gd(III)-based labels. We will present challenges in sample preparation and data analysis for functional and structural membrane protein studies using site-directed spin labeling techniques and give experimental details on EPR techniques providing information on side chain dynamics and water accessibility using nitroxide probes. An updated optimal Q-band DEER setup for nitroxide probes will be described, and its extension to gadolinium-containing samples will be addressed.
- Published
- 2019
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17. Dynamical decoupling of nitroxides in o-terphenyl: a study of temperature, deuteration and concentration effects.
- Author
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Soetbeer J, Hülsmann M, Godt A, Polyhach Y, and Jeschke G
- Abstract
We have characterized the temperature dependent transverse relaxation for 100 μM protonated and deuterated nitroxides in both protonated and deuterated o-terphenyl (OTP and dOTP) in distinct temperature regimes between 10 K and room temperature (RT). The choice of sample compositions allowed for a clear separation into slow and fast relaxation contributions based on a sum of two stretched exponential (SSE) parameterization between 10 and 60 K, and likewise at RT. The slow contribution is purely matrix dependent, while the fast process is determined by an interplay between a molecule and a matrix. Our systematic study of dynamical decoupling (DD) as a function of temperature (at 40, 80 K and RT), spin concentration, deuteration of nitroxide and/or OTP matrix and DD scheme for 1 to 5 refocusing pulses reveals that DD significantly prolongs phase memory times with respect to Hahn echo relaxation at 40 K, which we discuss in an SSE framework. At 80 K and RT, where (intra)molecular motions dominate relaxation, DD does not preserve electron spin coherence independent of the sample composition. Instead, we report a matrix nuclei dependent performance of the applied DD scheme at 40 K with Uhrig outperforming Carr-Purcell DD in OTP, and vice versa for a dOTP matrix.
- Published
- 2018
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18. Role of the nucleotidyl cyclase helical domain in catalytically active dimer formation.
- Author
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Vercellino I, Rezabkova L, Olieric V, Polyhach Y, Weinert T, Kammerer RA, Jeschke G, and Korkhov VM
- Subjects
- Adenylyl Cyclases genetics, Amino Acid Substitution, Bacterial Proteins chemistry, Bacterial Proteins genetics, Bacterial Proteins metabolism, Conserved Sequence, Crystallography, X-Ray, Cytosol enzymology, Dimerization, Enzyme Activation, Enzyme Stability, Guanylate Cyclase chemistry, Guanylate Cyclase genetics, Humans, Models, Molecular, Mutagenesis, Site-Directed, Mutant Proteins chemistry, Mutant Proteins genetics, Mutant Proteins metabolism, Mycobacterium avium Complex genetics, Receptors, Cell Surface chemistry, Receptors, Cell Surface genetics, Sequence Alignment, Sequence Analysis, Protein, Adenylyl Cyclases chemistry, Adenylyl Cyclases metabolism, Catalytic Domain, Mycobacterium avium Complex enzymology, Protein Conformation
- Abstract
Nucleotidyl cyclases, including membrane-integral and soluble adenylyl and guanylyl cyclases, are central components in a wide range of signaling pathways. These proteins are architecturally diverse, yet many of them share a conserved feature, a helical region that precedes the catalytic cyclase domain. The role of this region in cyclase dimerization has been a subject of debate. Although mutations within this region in various cyclases have been linked to genetic diseases, the molecular details of their effects on the enzymes remain unknown. Here, we report an X-ray structure of the cytosolic portion of the membrane-integral adenylyl cyclase Cya from Mycobacterium intracellulare in a nucleotide-bound state. The helical domains of each Cya monomer form a tight hairpin, bringing the two catalytic domains into an active dimerized state. Mutations in the helical domain of Cya mimic the disease-related mutations in human proteins, recapitulating the profiles of the corresponding mutated enzymes, adenylyl cyclase-5 and retinal guanylyl cyclase-1. Our experiments with full-length Cya and its cytosolic domain link the mutations to protein stability, and the ability to induce an active dimeric conformation of the catalytic domains. Sequence conservation indicates that this domain is an integral part of cyclase machinery across protein families and species. Our study provides evidence for a role of the helical domain in establishing a catalytically competent dimeric cyclase conformation. Our results also suggest that the disease-associated mutations in the corresponding regions of human nucleotidyl cyclases disrupt the normal helical domain structure., Competing Interests: The authors declare no conflict of interest., (Copyright © 2017 the Author(s). Published by PNAS.)
- Published
- 2017
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19. Solution structure of discoidal high-density lipoprotein particles with a shortened apolipoprotein A-I.
- Author
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Bibow S, Polyhach Y, Eichmann C, Chi CN, Kowal J, Albiez S, McLeod RA, Stahlberg H, Jeschke G, Güntert P, and Riek R
- Subjects
- Humans, Hydrophobic and Hydrophilic Interactions, Models, Molecular, Nuclear Magnetic Resonance, Biomolecular, Protein Conformation, alpha-Helical, Protein Structure, Secondary, Solutions, Apolipoprotein A-I chemistry, Lipoproteins, HDL chemistry
- Abstract
High-density lipoprotein (HDL) particles are cholesterol and lipid transport containers. Mature HDL particles destined for the liver develop through the formation of intermediate discoidal HDL particles, which are the primary acceptors for cholesterol. Here we present the three-dimensional structure of reconstituted discoidal HDL (rdHDL) particles, using a shortened construct of human apolipoprotein A-I, determined from a combination of nuclear magnetic resonance (NMR), electron paramagnetic resonance (EPR) and transmission electron microscopy (TEM) data. The rdHDL particles feature a protein double belt surrounding a lipid bilayer patch in an antiparallel fashion. The integrity of this structure is maintained by up to 28 salt bridges and a zipper-like pattern of cation-π interactions between helices 4 and 6. To accommodate a hydrophobic interior, a gross 'right-to-right' rotation of the helices after lipidation is necessary. The structure reflects the complexity required for a shuttling container to hold a fluid lipid or cholesterol interior at a protein:lipid ratio of 1:50.
- Published
- 2017
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20. Exploring the Strength of the H-Bond in Synthetic Models for Heme Proteins: The Importance of the N-H Acidity of the Distal Base.
- Author
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Alberti MN, Polyhach Y, Tzirakis MD, Tödtli L, Jeschke G, and Diederich F
- Subjects
- Heme-Binding Proteins, Hydrogen Bonding, Ligands, Models, Molecular, Spectrum Analysis, Carrier Proteins chemistry, Hemeproteins chemistry, Oxygen chemistry, Porphyrins chemistry
- Abstract
The distal hydrogen bond (H-bond) in dioxygen-binding proteins is crucial for the discrimination of O2 with respect to CO or NO. We report the preparation and characterization of a series of Zn(II) porphyrins, with one of three meso-phenyl rings bearing both an alkyl-tethered proximal imidazole ligand and a heterocyclic distal H-bond donor connected by a rigid acetylene spacer. Previously, we had validated the corresponding Co(II) complexes as synthetic model systems for dioxygen-binding heme proteins and demonstrated the structural requirements for proper distal H-bonding to Co(II) -bound dioxygen. Here, we systematically vary the H-bond donor ability of the distal heterocycles, as predicted based on pKa values. The H-bond in the dioxygen adducts of the Co(II) porphyrins was directly measured by Q-band Davies-ENDOR spectroscopy. It was shown that the strength of the hyperfine coupling between the dioxygen radical and the distal H-atom increases with enhanced acidity of the H-bond donor., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2016
- Full Text
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21. Glu-311 in External Loop 4 of the Sodium/Proline Transporter PutP Is Crucial for External Gate Closure.
- Author
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Bracher S, Guérin K, Polyhach Y, Jeschke G, Dittmer S, Frey S, Böhm M, and Jung H
- Subjects
- Amino Acid Sequence, Amino Acid Transport Systems, Neutral metabolism, Escherichia coli metabolism, Escherichia coli Proteins metabolism, Glutamine chemistry, Molecular Sequence Data, Protein Structure, Tertiary, Symporters metabolism, Amino Acid Transport Systems, Neutral chemistry, Escherichia coli Proteins chemistry, Molecular Dynamics Simulation, Symporters chemistry
- Abstract
The available structural information on LeuT and structurally related transporters suggests that external loop 4 (eL4) and the outer end of transmembrane domain (TM) 10' participate in the reversible occlusion of the outer pathway to the solute binding sites. Here, the functional significance of eL4 and the outer region of TM10' are explored using the sodium/proline symporter PutP as a model. Glu-311 at the tip of eL4, and various amino acids around the outer end of TM10' are identified as particularly crucial for function. Substitutions at these sites inhibit the transport cycle, and affect in part ligand binding. In addition, changes at selected sites induce a global structural alteration in the direction of an outward-open conformation. It is suggested that interactions between the tip of eL4 and the peptide backbone at the end of TM10' participate in coordinating conformational alterations underlying the alternating access mechanism of transport. Together with the structural information on LeuT-like transporters, the results further specify the idea that common design and functional principles are maintained across different transport families., (© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2016
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22. Modeling of the N-terminal Section and the Lumenal Loop of Trimeric Light Harvesting Complex II (LHCII) by Using EPR.
- Author
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Fehr N, Dietz C, Polyhach Y, von Hagens T, Jeschke G, and Paulsen H
- Subjects
- Models, Molecular, Protein Conformation, Biopolymers chemistry, Electron Spin Resonance Spectroscopy methods, Models, Chemical, Photosystem II Protein Complex chemistry
- Abstract
The major light harvesting complex II (LHCII) of green plants plays a key role in the absorption of sunlight, the regulation of photosynthesis, and in preventing photodamage by excess light. The latter two functions are thought to involve the lumenal loop and the N-terminal domain. Their structure and mobility in an aqueous environment are only partially known. Electron paramagnetic resonance (EPR) has been used to measure the structure of these hydrophilic protein domains in detergent-solubilized LHCII. A new technique is introduced to prepare LHCII trimers in which only one monomer is spin-labeled. These heterogeneous trimers allow to measure intra-molecular distances within one LHCII monomer in the context of a trimer by using double electron-electron resonance (DEER). These data together with data from electron spin echo envelope modulation (ESEEM) allowed to model the N-terminal protein section, which has not been resolved in current crystal structures, and the lumenal loop domain. The N-terminal domain covers only a restricted area above the superhelix in LHCII, which is consistent with the "Velcro" hypothesis to explain thylakoid grana stacking (Standfuss, J., van Terwisscha Scheltinga, A. C., Lamborghini, M., and Kühlbrandt, W. (2005) EMBO J. 24, 919-928). The conformation of the lumenal loop domain is surprisingly different between LHCII monomers and trimers but not between complexes with and without neoxanthin bound., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2015
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23. Extracellular loop 4 of the proline transporter PutP controls the periplasmic entrance to ligand binding sites.
- Author
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Raba M, Dunkel S, Hilger D, Lipiszko K, Polyhach Y, Jeschke G, Bracher S, Klare JP, Quick M, Jung H, and Steinhoff HJ
- Subjects
- Amino Acid Transport Systems, Neutral genetics, Binding Sites, Electron Spin Resonance Spectroscopy, Escherichia coli Proteins genetics, Hydrophobic and Hydrophilic Interactions, Ligands, Models, Molecular, Mutation, Phenylalanine chemistry, Protein Conformation, Symporters genetics, Amino Acid Transport Systems, Neutral chemistry, Amino Acid Transport Systems, Neutral metabolism, Escherichia coli Proteins chemistry, Escherichia coli Proteins metabolism, Symporters chemistry, Symporters metabolism
- Abstract
The Na(+)/proline symporter (PutP), like several other Na(+)-coupled symporters, belongs to the so-called LeuT-fold structural family, which features ten core transmembrane domains (cTMs) connected by extra- and intracellular loops. The role of these loops has been discussed in context with the gating function in the alternating access model of secondary active transport processes. Here we report the complete spin-labeling site scan of extracellular loop 4 (eL4) in PutP that reveals the presence of two α-helical segments, eL4a and eL4b. Among the eL4 residues that are directly implicated in the functional dynamics of the transporter, Phe314 in eL4b anchors the loop by means of hydrophobic contacts to cTM1 close to the ligand binding sites. We propose that ligand-induced conformational changes at the binding sites are transmitted via the anchoring residue to eL4 and through eL4 further to adjacent cTMs, leading to closure of the extracellular gate., (Copyright © 2014 Elsevier Ltd. All rights reserved.)
- Published
- 2014
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24. Suppression of ghost distances in multiple-spin double electron-electron resonance.
- Author
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von Hagens T, Polyhach Y, Sajid M, Godt A, and Jeschke G
- Subjects
- Electron Spin Resonance Spectroscopy, Electrons, Spin Labels, Models, Molecular
- Abstract
Distance measurements by pulse electron paramagnetic resonance techniques are increasingly applied to multiple-spin systems. In the double electron-electron resonance experiment, more than two dipolar coupled spins manifest in an increased total modulation depth and in sum and difference dipolar frequency contributions that give rise to additional peaks appearing in the distance distribution, which do not correspond to the real interspin distances of the system and are hence referred to as ghost contributions. These ghost contributions may be so prominent that they might be mistaken for real distance peaks or that real distance peaks shift their position or disappear. We present a simple approximate procedure to suppress ghost distances to a great extent by manipulating the experimentally obtained form factor during data analysis by a simple power scaling with a scaling exponent ζ(N) = 1/(1-N), with N being the number of coupled spins in the system. This approach requires neither further experimental effort nor exact knowledge about labelling and inversion efficiency. This should enable routine application to biological systems. The approach is validated on simulated test cases for up to five spins and applied to synthetic model samples. The suppression of ghost distances with the presented approach works best for symmetric geometries and rigid molecules which, at the same time, are the cases where ghost contributions are most disturbing. The distance distributions obtained by power scaling are consistent with distributions that were obtained with previously obtained alternative approaches and agree, in some cases, strikingly well with the expectations for the true interspin distance distributions.
- Published
- 2013
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- View/download PDF
25. EPR techniques to probe insertion and conformation of spin-labeled proteins in lipid bilayers.
- Author
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Bordignon E and Polyhach Y
- Subjects
- Electrons, Humans, Protein Structure, Secondary, Temperature, Water chemistry, bcl-2-Associated X Protein chemistry, bcl-2-Associated X Protein metabolism, Electron Spin Resonance Spectroscopy methods, Lipid Bilayers metabolism, Membrane Proteins metabolism, Molecular Conformation, Spin Labels
- Abstract
Electron paramagnetic resonance (EPR) spectroscopy of spin-labeled membrane proteins is a valuable biophysical technique to study structural details and conformational transitions of proteins close to their physiological environment, e.g., in liposomes, membrane bilayers, and nanodiscs. Unlike in nuclear magnetic resonance spectroscopy, having only one or few specific side chains labeled at a time with paramagnetic probes makes the size of the object under investigation irrelevant in terms of technique sensitivity. As a drawback, extensive site-directed mutagenesis is required in order to analyze the properties of the protein under investigation. EPR can provide detailed information on side chain dynamics of large membrane proteins or protein complexes embedded in membranes with an exquisite sensitivity for flexible regions and on water accessibility profiles across the membrane bilayer. Moreover, distances between the two spin-labeled side chains in membrane proteins can be detected with high precision in the 1.5-6 nm range at cryogenic temperatures. The application of EPR to membrane proteins still presents some challenges in terms of sample preparation, sensitivity, and data interpretation; thus no ready-to-go methodological recipes can be given. However this chapter describes the state of the art in the application of nitroxide-based site-directed spin labeling EPR to membrane proteins, with specific focus on the different types of information which can be obtained with continuous wave and pulsed techniques and on the challenges in sample preparation and data analysis for functional and structural membrane protein studies.
- Published
- 2013
- Full Text
- View/download PDF
26. High sensitivity and versatility of the DEER experiment on nitroxide radical pairs at Q-band frequencies.
- Author
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Polyhach Y, Bordignon E, Tschaggelar R, Gandra S, Godt A, and Jeschke G
- Subjects
- Electron Spin Resonance Spectroscopy, Liposomes chemistry, Spin Labels, Nitrogen Oxides chemistry
- Abstract
Measurement of distances with the Double Electron-Electron Resonance (DEER) experiment at X-band frequencies using a pair of nitroxides as spin labels is a popular biophysical tool for studying function-related conformational dynamics of proteins. The technique is intrinsically highly precise and can potentially access the range from 1.5 to 6-10 nm. However, DEER performance drops strongly when relaxation rates of the nitroxide spin labels are high and available material quantities are low, which is usually the case for membrane proteins reconstituted into liposomes. This leads to elevated noise levels, very long measurement times, reduced precision, and a decrease of the longest accessible distances. Here we quantify the performance improvement that can be achieved at Q-band frequencies (34.5 GHz) using a high-power spectrometer. More than an order of magnitude gain in sensitivity is obtained with a homebuilt setup equipped with a 150 W TWT amplifier by using oversized samples. The broadband excitation enabled by the high power ensures that orientation selection can be suppressed in most cases, which facilitates extraction of distance distributions. By varying pulse lengths, Q-band DEER can be switched between orientationally non-selective and selective regimes. Because of suppression of nuclear modulations from matrix protons and deuterons, analysis of the Q-band data is greatly simplified, particularly in cases of very small DEER modulation depth due to low binding affinity between proteins forming a complex or low labelling efficiency. Finally, we demonstrate that a commercial Q-band spectrometer can be readily adjusted to the high-power operation.
- Published
- 2012
- Full Text
- View/download PDF
27. Orientation selective DEER measurements on vinculin tail at X-band frequencies reveal spin label orientations.
- Author
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Abé C, Klose D, Dietrich F, Ziegler WH, Polyhach Y, Jeschke G, and Steinhoff HJ
- Subjects
- Algorithms, DNA, Complementary biosynthesis, DNA, Complementary genetics, Fourier Analysis, Nitrogen Oxides chemistry, Normal Distribution, Software, Solutions, Spin Labels, Electron Spin Resonance Spectroscopy methods, Vinculin chemistry
- Abstract
Double electron electron resonance (DEER) spectroscopy has been established as a valuable method to determine distances between spin labels bound to protein molecules. Caused by selective excitation of molecular orientations DEER primary data also depend on the mutual orientation of the spin labels. For a doubly spin labeled variant of the cytoskeletal protein vinculin tail strong orientation selection can be observed already at X-band frequencies, which allows us to reduce the problem to the relative orientation of two molecular axes and the spin-spin axis parameterized by three angles. A full grid search of parameter space reveals that the DEER experiment introduces parameter-space symmetry higher than the symmetry of the spin Hamiltonian. Thus, the number of equivalent parameter sets is twice as large as expected and the relative orientation of the two spin labels is ambiguous. Except for this inherent ambiguity the most probable relative orientation of the two spin labels can be determined with good confidence and moderate uncertainty by global fitting of a set of five DEER experiments at different offsets between pump and observer frequency. The experiment provides restraints on the angles between the z axis of the nitroxide molecular frame and the spin-spin vector and on the dihedral between the two z axes. When using the same type of label at both sites, assignment of the angle restraints is ambiguous and the sign of the dihedral restraint is also ambiguous., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
28. Rigid core and flexible terminus: structure of solubilized light-harvesting chlorophyll a/b complex (LHCII) measured by EPR.
- Author
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Dockter C, Müller AH, Dietz C, Volkov A, Polyhach Y, Jeschke G, and Paulsen H
- Subjects
- Chlorophyll chemistry, Chlorophyll A, Protein Structure, Quaternary, Protein Structure, Secondary, Protein Structure, Tertiary, Spin Labels, Light-Harvesting Protein Complexes chemistry, Pisum sativum enzymology
- Abstract
The structure of the major light-harvesting chlorophyll a/b complex (LHCII) was analyzed by pulsed EPR measurements and compared with the crystal structure. Site-specific spin labeling of the recombinant protein allowed the measurement of distance distributions over several intra- and intermolecular distances in monomeric and trimeric LHCII, yielding information on the protein structure and its local flexibility. A spin label rotamer library based on a molecular dynamics simulation was used to take the local mobility of spin labels into account. The core of LHCII in solution adopts a structure very similar or identical to the one seen in crystallized LHCII trimers with little motional freedom as indicated by narrow distance distributions along and between α helices. However, distances comprising the lumenal loop domain show broader distance distributions, indicating some mobility of this loop structure. Positions in the hydrophilic N-terminal domain, upstream of the first trans-membrane α helix, exhibit more and more mobility the closer they are to the N terminus. The nine amino acids at the very N terminus that have not been resolved in any of the crystal structure analyses give rise to very broad and possibly bimodal distance distributions, which may represent two families of preferred conformations.
- Published
- 2012
- Full Text
- View/download PDF
29. Rotamer libraries of spin labelled cysteines for protein studies.
- Author
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Polyhach Y, Bordignon E, and Jeschke G
- Subjects
- Cryoelectron Microscopy, Electron Spin Resonance Spectroscopy, Models, Molecular, Molecular Conformation, Cysteine chemistry, Muramidase chemistry, Peptide Fragments chemistry, Peptide Library, Spin Labels
- Abstract
Studies of structure and dynamics of proteins using site-directed spin labelling rely on explicit modelling of spin label conformations. The large computational effort associated with such modelling with molecular dynamics (MD) simulations can be avoided by a rotamer library approach based on a coarse-grained representation of the conformational space of the spin label. We show here that libraries of about 200 rotamers, obtained by iterative projection of a long MD trajectory of the free spin label onto a set of canonical dihedral angles, provide a representation of the underlying trajectory adequate for EPR distance measurements. Rotamer analysis was performed on selected X-ray structures of spin labelled T4 lysozyme mutants to characterize the spin label rotamer ensemble on a single protein site. Furthermore, predictions based on the rotamer library approach are shown to be in nearly quantitative agreement with electron paramagnetic resonance (EPR) distance data on the Na(+)/H(+) antiporter NhaA and on the light-harvesting complex LHCII whose structures are known from independent cryo electron microscopy and X-ray studies, respectively. Suggestions for the selection of labelling sites in proteins are given, limitations of the approach discussed, and requirements for further development are outlined.
- Published
- 2011
- Full Text
- View/download PDF
30. Refolding of the integral membrane protein light-harvesting complex II monitored by pulse EPR.
- Author
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Dockter C, Volkov A, Bauer C, Polyhach Y, Joly-Lopez Z, Jeschke G, and Paulsen H
- Subjects
- Apoproteins chemistry, Apoproteins metabolism, Electron Spin Resonance Spectroscopy, Electrons, Models, Biological, Mutation genetics, Protein Denaturation, Protein Structure, Secondary, Spin Labels, Time Factors, Light-Harvesting Protein Complexes chemistry, Light-Harvesting Protein Complexes metabolism, Membrane Proteins chemistry, Membrane Proteins metabolism, Pisum sativum metabolism, Protein Folding
- Abstract
The major light-harvesting chlorophyll a/b complex (LHCII) of the photosynthetic apparatus in plants self-organizes in vitro. The recombinant apoprotein, denatured in dodecyl sulfate, spontaneously folds when it is mixed with its pigments, chlorophylls, and carotenoids in detergent solution, and assembles into structurally authentic LHCII in the course of several minutes. Pulse EPR techniques, specifically double-electron-electron resonance (DEER), have been used to analyze protein folding during this process. Pairs of nitroxide labels were introduced site-specifically into recombinant LHCII and shown not to affect the stability and function of the pigment-protein complex. Interspin distance distributions between two spin pairs were measured at various time points, one pair located on either end of the second transmembrane helix (helix 3), the other one located near the luminal ends of the intertwined transmembrane helices 1 and 4. In the dodecyl sulfate-solubilized apoprotein, both distance distributions were consistent with a random-coil protein structure. A rapid freeze-quench experiment on the latter spin pair indicated that 1 s after initiating reconstitution the protein structure is virtually unchanged. Subsequently, both distance distributions monitored protein folding in the same time range in which the assembly of chlorophylls into the complex had been observed. The positioning of the spin pair spanning the hydrophobic core of LHCII clearly preceded the juxtaposition of the spin pair on the luminal side of the complex. This indicates that superhelix formation of helices 1 and 4 is a late step in LHCII assembly.
- Published
- 2009
- Full Text
- View/download PDF
31. Transmembrane signaling in the maltose ABC transporter MalFGK2-E: periplasmic MalF-P2 loop communicates substrate availability to the ATP-bound MalK dimer.
- Author
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Grote M, Polyhach Y, Jeschke G, Steinhoff HJ, Schneider E, and Bordignon E
- Subjects
- ATP-Binding Cassette Transporters chemistry, ATP-Binding Cassette Transporters genetics, Dimerization, Electron Spin Resonance Spectroscopy, Escherichia coli genetics, Escherichia coli metabolism, Escherichia coli Proteins chemistry, Escherichia coli Proteins genetics, Maltose metabolism, Models, Molecular, Monosaccharide Transport Proteins chemistry, Monosaccharide Transport Proteins genetics, Mutagenesis, Site-Directed, Periplasm, Protein Conformation, Signal Transduction, ATP-Binding Cassette Transporters metabolism, Adenosine Triphosphate metabolism, Cell Membrane metabolism, Escherichia coli Proteins metabolism, Monosaccharide Transport Proteins metabolism
- Abstract
ABC transporters are ubiquitous membrane proteins that translocate solutes across biological membranes at the expense of ATP. In prokaryotic ABC importers, the extracytoplasmic anchoring of the substrate-binding protein (receptor) is emerging as a key determinant for the structural rearrangements in the cytoplasmically exposed ATP-binding cassette domains and in the transmembrane gates during the nucleotide cycle. Here the molecular mechanism of such signaling events was addressed by electron paramagnetic resonance spectroscopy of spin-labeled ATP-binding cassette maltose transporter variants (MalFGK2-E). A series of doubly spin-labeled mutants in the MalF-P2 domain involving positions 92, 205, 239, 252, and 273 and one triple mutant labeled at positions 205/252 in P2 and 83 in the Q-loop of MalK were assayed. The EPR data revealed that the substrate-binding protein MalE is bound to the transporter throughout the transport cycle. Concomitantly with the three conformations of the ATP-binding cassette MalK2, three functionally relevant conformations are found also in the periplasmic MalF-P2 loop, strictly dependent on cytoplasmic nucleotide binding and periplasmic docking of liganded MalE to MalFG. The reciprocal communication across the membrane unveiled here gives first insights into the stimulatory effect of MalE on the ATPase activity, and it is suggested to be an important mechanistic feature of receptor-coupled ABC transporters.
- Published
- 2009
- Full Text
- View/download PDF
32. Backbone structure of transmembrane domain IX of the Na+/proline transporter PutP of Escherichia coli.
- Author
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Hilger D, Polyhach Y, Jung H, and Jeschke G
- Subjects
- Amino Acid Sequence, Amino Acid Transport Systems, Neutral genetics, Electron Spin Resonance Spectroscopy methods, Escherichia coli, Escherichia coli Proteins genetics, Models, Chemical, Protein Conformation, Protein Structure, Secondary, Symporters genetics, Amino Acid Transport Systems, Neutral chemistry, Escherichia coli Proteins chemistry, Symporters chemistry
- Abstract
The backbone structure is determined by site-directed spin labeling, double electron electron resonance measurements of distances, and modeling in terms of a helix-loop-helix construct for a transmembrane domain that is supposed to line the translocation pathway in the 54.3 kDa Na(+)/proline symporter PutP of Escherichia coli. The conformational distribution of the spin labels is accounted for by a rotamer library. An ensemble of backbone models with a root mean-square deviation of less than 2 A is obtained. These models exhibit a pronounced kink near residue T341, which is involved in substrate binding. The kink may be associated with a hinge that allows the protein to open and close an inwardly oriented cavity.
- Published
- 2009
- Full Text
- View/download PDF
33. A comparative electron paramagnetic resonance study of the nucleotide-binding domains' catalytic cycle in the assembled maltose ATP-binding cassette importer.
- Author
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Grote M, Bordignon E, Polyhach Y, Jeschke G, Steinhoff HJ, and Schneider E
- Subjects
- ATP-Binding Cassette Transporters genetics, Alkanesulfonates metabolism, Biomechanical Phenomena, Catalysis, Computational Biology, Cross-Linking Reagents metabolism, Crystallography, X-Ray, Cysteine, Dimerization, Electron Spin Resonance Spectroscopy, Monosaccharide Transport Proteins genetics, Mutation, Protein Structure, Quaternary, Protein Structure, Tertiary, Solubility, Spin Labels, ATP-Binding Cassette Transporters chemistry, ATP-Binding Cassette Transporters metabolism, Monosaccharide Transport Proteins chemistry, Monosaccharide Transport Proteins metabolism, Nucleotides metabolism
- Abstract
We present a quantitative analysis of conformational changes of the nucleotide-binding subunits, MalK(2), of the maltose ATP-binding cassette importer MalFGK(2) during the transport cycle. Distance changes occurring between selected residues were monitored in the full transporter by site-directed spin-labeling electron paramagnetic resonance spectroscopy and site-directed chemical cross-linking. We considered S83C and A85C from the conserved Q-loop and V117C located on the outer surface of MalK. Additionally, two native cysteines (C350, C360) were included in the study. On ATP binding, small rearrangements between the native sites, and no distance changes between positions 117 were detected. In contrast, positions 85 come closer together in the ATP-bound state and in the vanadate-trapped intermediate and move back toward the apo-state after ATP hydrolysis. The distance between positions 83 is shown to slightly decrease on ATP binding, and to further decrease after ATP hydrolysis. Results from cross-linking experiments are in agreement with these findings. The data are compared with in silico spin-labeled x-ray structures from both isolated MalK(2) and the MalFGK(2)-E complex. Our results are consistent with a slightly modified "tweezers-like" model of closure and reopening of MalK(2) during the catalytic cycle, and show an unforeseen potential interaction between MalK and the transmembrane subunit MalG.
- Published
- 2008
- Full Text
- View/download PDF
34. Distance measurements on spin-labelled biomacromolecules by pulsed electron paramagnetic resonance.
- Author
-
Jeschke G and Polyhach Y
- Subjects
- Sensitivity and Specificity, Temperature, Electron Spin Resonance Spectroscopy, Macromolecular Substances chemistry, Spin Labels
- Abstract
The biological function of protein, DNA, and RNA molecules often depends on relative movements of domains with dimensions of a few nanometers. This length scale can be accessed by distance measurements between spin labels if pulsed electron paramagnetic resonance (EPR) techniques such as electron-electron double resonance (ELDOR) and double-quantum EPR are used. The approach does not require crystalline samples and is well suited to biomacromolecules with an intrinsic flexibility as distributions of distances can be measured. Furthermore, oligomerization or complexation of biomacromolecules can also be studied, even if it is incomplete. The sensitivity of the technique and the reliability of the measured distance distribution depend on careful optimization of the experimental conditions and procedures for data analysis. Interpretation of spin-to-spin distance distributions in terms of the structure of the biomacromolecules furthermore requires a model for the conformational distribution of the spin labels.
- Published
- 2007
- Full Text
- View/download PDF
35. Spin pair geometry revealed by high-field DEER in the presence of conformational distributions.
- Author
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Polyhach Y, Godt A, Bauer C, and Jeschke G
- Subjects
- Computer Simulation, Molecular Conformation, Spin Labels, Algorithms, Biopolymers chemistry, Magnetic Resonance Spectroscopy methods, Models, Chemical, Models, Molecular
- Abstract
Orientation selection on two nitroxide-labelled shape-persistent molecules is demonstrated by high-field pulsed electron-electron double resonance experiments at a frequency of 95 GHz with a commercial spectrometer. The experiments are performed with fixed observer and pump frequencies by variation of the magnetic field, so that the variation of both the dipolar frequencies and the modulation depths can be analyzed. By applying the deadtime-free four-pulse double electron-electron resonance (DEER) sequence, the lineshapes of the dipolar spectra are obtained. In the investigated linear biradical and equilateral triradical the nitroxide labels undergo restricted dynamics, so that their relative orientations are not fixed, but are correlated to some extent. In this situation, the general dependence of the dipolar spectra on the observer field can be satisfyingly modelled by simple geometrical models that involve only one rotational degree of freedom for the biradical and two rotational degrees of freedom for the triradical. A somewhat better agreement of the dipolar lineshapes for the biradical is obtained by simulations based on a molecular dynamics trajectory. For the triradical, small but significant deviations of the lineshape are observed with both models, indicating that the technique can reveal deficiencies in modelling of the conformational ensemble of a macromolecule.
- Published
- 2007
- Full Text
- View/download PDF
36. EPR and NMR in powders of doped and undoped IV-VI crystals.
- Author
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Zayachuk D, Polyhach Y, Slynko E, Khandozhko O, and Rudowicz C
- Subjects
- Crystallization, Lead chemistry, Semiconductors, Tellurium chemistry, Temperature, Time Factors, Tin chemistry, Electron Spin Resonance Spectroscopy, Gadolinium chemistry, Magnetic Resonance Spectroscopy, Powders chemistry
- Abstract
The results of the extensive investigations of the variation of the EPR and NMR spectra of active centers due to the existence of the native defects generated by disorder in the IV-VI semiconductor matrices are presented. Both undoped and doped with Gd impurity powder samples of different grain sizes, made from Pb(1-x)Sn(x)Te crystals with the composition in the range 0 < or = x < or = 0.2 were studied. Impurity Gd ions were used as the paramagnetic EPR probe, whereas the 207 Pb nuclei as the NMR probe. The following aspects have been ascertained. (i) Grinding of the initial single crystals into powders leads to an additional component line appearing in the NMR spectra of the 207 Pb nuclei and also to a significant increase in the intensity of EPR spectra of the impurity Gd ions. (ii) Both the Gd EPR spectra as well as the 207 Pb NMR spectra undergo modifications due to isothermal annealing, whereas the character of these modifications is determined by both the temperature and duration of the thermal treatment applied. (iii) Some characteristic correlation between the variation of the EPR spectra of impurity Gd ions and that of the NMR spectra of 207 Pb nuclei, which results from the annealing of the samples, has been observed. Experimental results are interpreted based on the prevailing models of the behavior of the doped impurities and the native defects in the lead and tin telluride crystals.
- Published
- 2004
- Full Text
- View/download PDF
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