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

MPS-Authors
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Grohe,  K.
Research Group of Solid-State NMR-2, MPI for Biophysical Chemistry, Max Planck Society;

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Vasa,  S. K.
Research Group of Solid-State NMR-2, MPI for Biophysical Chemistry, Max Planck Society;

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Giller,  K.
Department of NMR-Based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Becker,  S.
Department of NMR-Based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

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Linser,  R.
Research Group of Solid-State NMR-2, MPI for Biophysical Chemistry, Max Planck Society;

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Citation

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.


Cite as: http://hdl.handle.net/11858/00-001M-0000-002D-3134-A
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.