[NMR paper] Beyond Structural Biology to Functional Biology: Solid-State NMR Experiments and Strategies for Understanding the M2 Proton Channel Conductance.
Related ArticlesBeyond Structural Biology to Functional Biology: Solid-State NMR Experiments and Strategies for Understanding the M2 Proton Channel Conductance.
J Phys Chem B. 2017 Apr 20;:
Authors: Qin H, Miao Y, Cross TA, Fu R
Abstract
In terms of structural biology, solid-state NMR experiments and strategies have been well established for resonance assignments leading to the determination of three-dimensional structures of insoluble membrane proteins in their native-like environment. It is also known that NMR has the unique capabilities to characterize structure-function relationships of membrane-bound biological systems beyond structural biology. Here, we report on solid-state NMR experiments and strategies for extracting functional activities on a sub-msec time scale. Specifically, we use the His37-labeled full length M2 (M2FL) protein of Influenza A virus embedded in synthetic lipid bilayers as an example to characterize the proton conduction mechanism and kinetics. The integral membrane M2 protein assembles as a tetrameric bundle to form a proton-conducting channel that is activated by low pH and is essential for the viral lifecycle. Our results present convincing evidence for the formation of imidazolium-imidazole hydrogen-bonds in the His37 tetrad at low pH and that these hydrogen bonds have a low barrier that facilitates the proton conduction mechanism in the M2FL protein. Moreover, it has been possible to measure hydronium ion exchange between water and the protons in the His37 NH bonds based on chemical exchange spectroscopy with minimized spin diffusion. The results identify an exchange rate constant of ~4000 s-1 for pH 5.8 at -10°C.
PMID: 28425709 [PubMed - as supplied by publisher]
Structural biology applications of solid state MAS DNP NMR #DNPNMR
From The DNP-NMR Blog:
Structural biology applications of solid state MAS DNP NMR #DNPNMR
Akbey, U. and H. Oschkinat, Structural biology applications of solid state MAS DNP NMR. J Magn Reson, 2016. 269: p. 213-24.
http://www.ncbi.nlm.nih.gov/pubmed/27095695
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08-31-2016 02:34 PM
[NMR paper] Solid-State NMR Investigation of the Conformation, Proton Conduction, and Hydration of the Influenza B Virus M2 Transmembrane Proton Channel.
Solid-State NMR Investigation of the Conformation, Proton Conduction, and Hydration of the Influenza B Virus M2 Transmembrane Proton Channel.
Related Articles Solid-State NMR Investigation of the Conformation, Proton Conduction, and Hydration of the Influenza B Virus M2 Transmembrane Proton Channel.
J Am Chem Soc. 2016 Jun 10;
Authors: Williams JK, Tietze D, Lee M, Wang J, Hong M
Abstract
Together with the influenza A virus, influenza B virus causes seasonal flu epidemics. The M2 protein of influenza B (BM2) forms a tetrameric...
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06-11-2016 01:09 PM
[NMR paper] Solid-state NMR: An emerging technique in structural biology of self-assemblies.
Solid-state NMR: An emerging technique in structural biology of self-assemblies.
Related Articles Solid-state NMR: An emerging technique in structural biology of self-assemblies.
Biophys Chem. 2015 Jul 16;
Authors: Habenstein B, Loquet A
Abstract
Protein self-assemblies are ubiquitous biological systems involved in many cellular processes, ranging from bacterial and viral infection to the propagation of neurodegenerative disorders. Studying the atomic three-dimensional structures of protein self-assemblies is a particularly...
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08-04-2015 03:00 PM
Solid-state NMR enhanced by dynamic nuclear polarization as a novel tool for ribosome structural biology
From The DNP-NMR Blog:
Solid-state NMR enhanced by dynamic nuclear polarization as a novel tool for ribosome structural biology
Gelis, I., et al., Solid-state NMR enhanced by dynamic nuclear polarization as a novel tool for ribosome structural biology. J. Biomol. NMR, 2013. 56(2): p. 85-93.
http://dx.doi.org/10.1007/s10858-013-9721-2
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09-09-2013 05:14 PM
Heat management strategies for solid-state NMR of functional proteins
Heat management strategies for solid-state NMR of functional proteins
September 2012
Publication year: 2012
Source:Journal of Magnetic Resonance, Volume 222</br>
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Modern solid-state NMR methods can acquire high-resolution protein spectra for structure determination. However, these methods use rapid sample spinning and intense decoupling fields that can heat and denature the protein being studied. Here we present a strategy to avoid destroying valuable samples. We advocate first creating a sacrificial sample, which contains unlabeled protein (or no protein) in buffer...
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02-03-2013 10:13 AM
Heat Management Strategies for Solid-state NMR of Functional Proteins
Heat Management Strategies for Solid-state NMR of Functional Proteins
Publication year: 2012
Source:Journal of Magnetic Resonance</br>
Daniel J. Fowler, Michael J. Harris, Lynmarie K. Thompson</br>
Modern solid-state NMR methods can acquire high-resolution protein spectra for structure determination. However, these methods use rapid sample spinning and intense decoupling fields that can heat and denature the protein being studied. Here we present a strategy to avoid destroying valuable samples. We advocate first creating a sacrificial sample, which contains unlabeled...
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07-14-2012 01:53 PM
Structural topology of phospholamban pentamer in lipid bilayers by a hybrid solution and solid-state NMR method [Biophysics and Computational Biology]
Structural topology of phospholamban pentamer in lipid bilayers by a hybrid solution and solid-state NMR method
Verardi, R., Shi, L., Traaseth, N. J., Walsh, N., Veglia, G....
Date: 2011-05-31
Phospholamban (PLN) is a type II membrane protein that inhibits the sarcoplasmic reticulum Ca2+-ATPase (SERCA), thereby regulating calcium homeostasis in cardiac muscle. In membranes, PLN forms pentamers that have been proposed to function either as a storage for active monomers or as ion channels. Here, we report the T-state structure of pentameric PLN solved by a hybrid solution and...
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05-31-2011 11:41 PM
Solid-state 2H NMR relaxation illuminates functional dynamics of retinal cofactor in membrane activation of rhodopsin [Biophysics and Computational Biology]
Solid-state 2H NMR relaxation illuminates functional dynamics of retinal cofactor in membrane activation of rhodopsin
Struts, A. V., Salgado, G. F. J., Brown, M. F....
Date: 2011-05-17
Rhodopsin is a canonical member of the family of G protein-coupled receptors, which transmit signals across cellular membranes and are linked to many drug interventions in humans. Here we show that solid-state 2H NMR relaxation allows investigation of light-induced changes in local ps–ns time scale motions of retinal bound to rhodopsin. Site-specific 2H labels were introduced into methyl groups of the...
Beyond Structural Biology to Functional Biology: Solid-State NMR Experiments and Strategies for Understanding the M2 Proton Channel Conductance.
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