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Site-Resolved Measurement of Microsecond-to-Millisecond Conformational-Exchange Processes in Proteins by Solid-State NMR Spectroscopy

[Image: see text] We demonstrate that conformational exchange processes in proteins on microsecond-to-millisecond time scales can be detected and quantified by solid-state NMR spectroscopy. We show two independent approaches that measure the effect of conformational exchange on transverse relaxation...

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Autores principales: Tollinger, Martin, Sivertsen, Astrid C., Meier, Beat H., Ernst, Matthias, Schanda, Paul
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2012
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3557925/
https://www.ncbi.nlm.nih.gov/pubmed/22908968
http://dx.doi.org/10.1021/ja303591y
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author Tollinger, Martin
Sivertsen, Astrid C.
Meier, Beat H.
Ernst, Matthias
Schanda, Paul
author_facet Tollinger, Martin
Sivertsen, Astrid C.
Meier, Beat H.
Ernst, Matthias
Schanda, Paul
author_sort Tollinger, Martin
collection PubMed
description [Image: see text] We demonstrate that conformational exchange processes in proteins on microsecond-to-millisecond time scales can be detected and quantified by solid-state NMR spectroscopy. We show two independent approaches that measure the effect of conformational exchange on transverse relaxation parameters, namely Carr–Purcell–Meiboom–Gill relaxation-dispersion experiments and measurement of differential multiple-quantum coherence decay. Long coherence lifetimes, as required for these experiments, are achieved by the use of highly deuterated samples and fast magic-angle spinning. The usefulness of the approaches is demonstrated by application to microcrystalline ubiquitin. We detect a conformational exchange process in a region of the protein for which dynamics have also been observed in solution. Interestingly, quantitative analysis of the data reveals that the exchange process is more than 1 order of magnitude slower than in solution, and this points to the impact of the crystalline environment on free energy barriers.
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spelling pubmed-35579252013-01-31 Site-Resolved Measurement of Microsecond-to-Millisecond Conformational-Exchange Processes in Proteins by Solid-State NMR Spectroscopy Tollinger, Martin Sivertsen, Astrid C. Meier, Beat H. Ernst, Matthias Schanda, Paul J Am Chem Soc [Image: see text] We demonstrate that conformational exchange processes in proteins on microsecond-to-millisecond time scales can be detected and quantified by solid-state NMR spectroscopy. We show two independent approaches that measure the effect of conformational exchange on transverse relaxation parameters, namely Carr–Purcell–Meiboom–Gill relaxation-dispersion experiments and measurement of differential multiple-quantum coherence decay. Long coherence lifetimes, as required for these experiments, are achieved by the use of highly deuterated samples and fast magic-angle spinning. The usefulness of the approaches is demonstrated by application to microcrystalline ubiquitin. We detect a conformational exchange process in a region of the protein for which dynamics have also been observed in solution. Interestingly, quantitative analysis of the data reveals that the exchange process is more than 1 order of magnitude slower than in solution, and this points to the impact of the crystalline environment on free energy barriers. American Chemical Society 2012-08-21 2012-09-12 /pmc/articles/PMC3557925/ /pubmed/22908968 http://dx.doi.org/10.1021/ja303591y Text en Copyright © 2012 American Chemical Society Terms of Use (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html)
spellingShingle Tollinger, Martin
Sivertsen, Astrid C.
Meier, Beat H.
Ernst, Matthias
Schanda, Paul
Site-Resolved Measurement of Microsecond-to-Millisecond Conformational-Exchange Processes in Proteins by Solid-State NMR Spectroscopy
title Site-Resolved Measurement of Microsecond-to-Millisecond Conformational-Exchange Processes in Proteins by Solid-State NMR Spectroscopy
title_full Site-Resolved Measurement of Microsecond-to-Millisecond Conformational-Exchange Processes in Proteins by Solid-State NMR Spectroscopy
title_fullStr Site-Resolved Measurement of Microsecond-to-Millisecond Conformational-Exchange Processes in Proteins by Solid-State NMR Spectroscopy
title_full_unstemmed Site-Resolved Measurement of Microsecond-to-Millisecond Conformational-Exchange Processes in Proteins by Solid-State NMR Spectroscopy
title_short Site-Resolved Measurement of Microsecond-to-Millisecond Conformational-Exchange Processes in Proteins by Solid-State NMR Spectroscopy
title_sort site-resolved measurement of microsecond-to-millisecond conformational-exchange processes in proteins by solid-state nmr spectroscopy
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3557925/
https://www.ncbi.nlm.nih.gov/pubmed/22908968
http://dx.doi.org/10.1021/ja303591y
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