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  Non-equilibrium hydrogen exchange for determination of H-bond strength and water accessibility in solid proteins.

Grohe, K., Movellan, K. T., Vasa, S. K., Giller, K., Becker, S., & Linser, R. (2017). Non-equilibrium hydrogen exchange for determination of H-bond strength and water accessibility in solid proteins. Journal of Biomolecular NMR, 68(1), 7-17.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002D-3134-A Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002D-61B6-5
Genre: Journal Article

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 Creators:
Grohe, K.1, Author              
Movellan, K. T., Author
Vasa, S. K.1, Author              
Giller, K.2, Author              
Becker, S.2, Author              
Linser, R.1, Author              
Affiliations:
1Research Group of Solid-State NMR-2, MPI for Biophysical Chemistry, Max Planck Society, ou_1950286              
2Department of NMR-Based Structural Biology, MPI for biophysical chemistry, Max Planck Society, ou_578567              

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Free keywords: Solid-state NMR, H-bond determination, Water accessibility, Protein structure, Proton detection, MAS
 Abstract: We demonstrate measurement of non-equilibrium backbone amide hydrogen-deuterium exchange rates (HDX) for solid proteins. The target of this study are the slowly exchanging residues in solid samples, which are associated with stable secondary-structural elements of proteins. These hydrogen exchange processes escape methods measuring equilibrium exchange rates of faster processes. The method was applied to a micro-crystalline preparation of the SH3 domain of chicken α-spectrin. Therefore, from a 100% back-exchanged micro-crystalline protein preparation, the supernatant buffer was exchanged by a partially deuterated buffer to reach a final protonation level of approximately 20% before packing the sample in a 1.3 mm rotor. Tracking of the HN peak intensities for 2 weeks reports on site-specific hydrogen bond strength and also likely reflects water accessibility in a qualitative manner. H/D exchange can be directly determined for hydrogen-bonded amides using 1H detection under fast magic angle spinning. This approach complements existing methods and provides the means to elucidate interesting site-specific characteristics for protein functionality in the solid state.

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Language(s): eng - English
 Dates: 2017-04-092017-05
 Publication Status: Published in print
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Title: Journal of Biomolecular NMR
Source Genre: Journal
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Pages: - Volume / Issue: 68 (1) Sequence Number: - Start / End Page: 7 - 17 Identifier: -