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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2012
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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. |
format | Online Article Text |
id | pubmed-3557925 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
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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