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Characterization of Protein–Protein Interfaces in Large Complexes by Solid-State NMR Solvent Paramagnetic Relaxation Enhancements
[Image: see text] Solid-state NMR is becoming a viable alternative for obtaining information about structures and dynamics of large biomolecular complexes, including ones that are not accessible to other high-resolution biophysical techniques. In this context, methods for probing protein–protein int...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2017
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5590091/ https://www.ncbi.nlm.nih.gov/pubmed/28780861 http://dx.doi.org/10.1021/jacs.7b03875 |
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author | Öster, Carl Kosol, Simone Hartlmüller, Christoph Lamley, Jonathan M. Iuga, Dinu Oss, Andres Org, Mai-Liis Vanatalu, Kalju Samoson, Ago Madl, Tobias Lewandowski, Józef R. |
author_facet | Öster, Carl Kosol, Simone Hartlmüller, Christoph Lamley, Jonathan M. Iuga, Dinu Oss, Andres Org, Mai-Liis Vanatalu, Kalju Samoson, Ago Madl, Tobias Lewandowski, Józef R. |
author_sort | Öster, Carl |
collection | PubMed |
description | [Image: see text] Solid-state NMR is becoming a viable alternative for obtaining information about structures and dynamics of large biomolecular complexes, including ones that are not accessible to other high-resolution biophysical techniques. In this context, methods for probing protein–protein interfaces at atomic resolution are highly desirable. Solvent paramagnetic relaxation enhancements (sPREs) proved to be a powerful method for probing protein–protein interfaces in large complexes in solution but have not been employed toward this goal in the solid state. We demonstrate that (1)H and (15)N relaxation-based sPREs provide a powerful tool for characterizing intermolecular interactions in large assemblies in the solid state. We present approaches for measuring sPREs in practically the entire range of magic angle spinning frequencies used for biomolecular studies and discuss their benefits and limitations. We validate the approach on crystalline GB1, with our experimental results in good agreement with theoretical predictions. Finally, we use sPREs to characterize protein–protein interfaces in the GB1 complex with immunoglobulin G (IgG). Our results suggest the potential existence of an additional binding site and provide new insights into GB1:IgG complex structure that amend and revise the current model available from studies with IgG fragments. We demonstrate sPREs as a practical, widely applicable, robust, and very sensitive technique for determining intermolecular interaction interfaces in large biomolecular complexes in the solid state. |
format | Online Article Text |
id | pubmed-5590091 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-55900912017-09-11 Characterization of Protein–Protein Interfaces in Large Complexes by Solid-State NMR Solvent Paramagnetic Relaxation Enhancements Öster, Carl Kosol, Simone Hartlmüller, Christoph Lamley, Jonathan M. Iuga, Dinu Oss, Andres Org, Mai-Liis Vanatalu, Kalju Samoson, Ago Madl, Tobias Lewandowski, Józef R. J Am Chem Soc [Image: see text] Solid-state NMR is becoming a viable alternative for obtaining information about structures and dynamics of large biomolecular complexes, including ones that are not accessible to other high-resolution biophysical techniques. In this context, methods for probing protein–protein interfaces at atomic resolution are highly desirable. Solvent paramagnetic relaxation enhancements (sPREs) proved to be a powerful method for probing protein–protein interfaces in large complexes in solution but have not been employed toward this goal in the solid state. We demonstrate that (1)H and (15)N relaxation-based sPREs provide a powerful tool for characterizing intermolecular interactions in large assemblies in the solid state. We present approaches for measuring sPREs in practically the entire range of magic angle spinning frequencies used for biomolecular studies and discuss their benefits and limitations. We validate the approach on crystalline GB1, with our experimental results in good agreement with theoretical predictions. Finally, we use sPREs to characterize protein–protein interfaces in the GB1 complex with immunoglobulin G (IgG). Our results suggest the potential existence of an additional binding site and provide new insights into GB1:IgG complex structure that amend and revise the current model available from studies with IgG fragments. We demonstrate sPREs as a practical, widely applicable, robust, and very sensitive technique for determining intermolecular interaction interfaces in large biomolecular complexes in the solid state. American Chemical Society 2017-08-07 2017-09-06 /pmc/articles/PMC5590091/ /pubmed/28780861 http://dx.doi.org/10.1021/jacs.7b03875 Text en Copyright © 2017 American Chemical Society This is an open access article published under a Creative Commons Attribution (CC-BY) License (http://pubs.acs.org/page/policy/authorchoice_ccby_termsofuse.html) , which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited. |
spellingShingle | Öster, Carl Kosol, Simone Hartlmüller, Christoph Lamley, Jonathan M. Iuga, Dinu Oss, Andres Org, Mai-Liis Vanatalu, Kalju Samoson, Ago Madl, Tobias Lewandowski, Józef R. Characterization of Protein–Protein Interfaces in Large Complexes by Solid-State NMR Solvent Paramagnetic Relaxation Enhancements |
title | Characterization
of Protein–Protein Interfaces
in Large Complexes by Solid-State NMR Solvent Paramagnetic Relaxation
Enhancements |
title_full | Characterization
of Protein–Protein Interfaces
in Large Complexes by Solid-State NMR Solvent Paramagnetic Relaxation
Enhancements |
title_fullStr | Characterization
of Protein–Protein Interfaces
in Large Complexes by Solid-State NMR Solvent Paramagnetic Relaxation
Enhancements |
title_full_unstemmed | Characterization
of Protein–Protein Interfaces
in Large Complexes by Solid-State NMR Solvent Paramagnetic Relaxation
Enhancements |
title_short | Characterization
of Protein–Protein Interfaces
in Large Complexes by Solid-State NMR Solvent Paramagnetic Relaxation
Enhancements |
title_sort | characterization
of protein–protein interfaces
in large complexes by solid-state nmr solvent paramagnetic relaxation
enhancements |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5590091/ https://www.ncbi.nlm.nih.gov/pubmed/28780861 http://dx.doi.org/10.1021/jacs.7b03875 |
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