PuSH - Publikationsserver des Helmholtz Zentrums München

Lopez del Amo, J.M.* ; Fink, U.* ; Reif, B.

Quantification of protein backbone hydrogen-deuterium exchange rates by solid state NMR spectroscopy.

J. Biomol. NMR 48, 203-212 (2010)
DOI Verlagsversion bestellen
Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
We present the quantification of backbone amide hydrogen-deuterium exchange rates (HDX) for immobilized proteins. The experiments make use of the deuterium isotope effect on the amide nitrogen chemical shift, as well as on proton dilution by deuteration. We find that backbone amides in the microcrystalline α-spectrin SH3 domain exchange rather slowly with the solvent (with exchange rates negligible within the individual (15)N-T (1) timescales). We observed chemical exchange for 6 residues with HDX exchange rates in the range from 0.2 to 5 s(-1). Backbone amide (15)N longitudinal relaxation times that we determined previously are not significantly affected for most residues, yielding no systematic artifacts upon quantification of backbone dynamics (Chevelkov et al. 2008b). Significant exchange was observed for the backbone amides of R21, S36 and K60, as well as for the sidechain amides of N38, N35 and for W41ε. These residues could not be fit in our previous motional analysis, demonstrating that amide proton chemical exchange needs to be considered in the analysis of protein dynamics in the solid-state, in case D(2)O is employed as a solvent for sample preparation. Due to the intrinsically long (15)N relaxation times in the solid-state, the approach proposed here can expand the range of accessible HDX rates in the intermediate regime that is not accessible so far with exchange quench and MEXICO type experiments.
Altmetric
Weitere Metriken?
Zusatzinfos bearbeiten [➜Einloggen]
Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter MAS solid-state NMR; Hydrogen-deuterium exchange; Relaxation; Perdeuterated proteins; Alpha-spectrin SH3
ISSN (print) / ISBN 0925-2738
e-ISSN 1573-5001
Quellenangaben Band: 48, Heft: 4, Seiten: 203-212 Artikelnummer: , Supplement: ,
Verlag Springer
Begutachtungsstatus Peer reviewed