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Hydrogen‐deuterium exchange in membrane proteins monitored by IR spectroscopy: A new tool to resolve protein structure and dynamics
Author(s) -
Vigano C.,
Smeyers M.,
Raussens V.,
Scheirlinckx F.,
Ruysschaert J.M.,
Goormaghtigh E.
Publication year - 2004
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.20035
Subject(s) - chemistry , hydrogen–deuterium exchange , kinetics , fourier transform infrared spectroscopy , deuterium , spectroscopy , protein secondary structure , protein dynamics , protein structure , membrane , infrared spectroscopy , molecular dynamics , attenuated total reflection , amide , mass spectrometry , computational chemistry , chromatography , biochemistry , chemical engineering , organic chemistry , physics , quantum mechanics , engineering
As more and more high‐resolution structures of proteins become available, the new challenge is the understanding of these small conformational changes that are responsible for protein activity. Specialized difference Fourier transform infrared (FTIR) techniques allow the recording of side‐chain modifications or minute secondary structure changes. Yet, large domain movements remain usually unnoticed. FTIR spectroscopy provides a unique opportunity to record 1 H/ 2 H exchange kinetics at the level of the amide proton. This approach is extremely sensitive to tertiary structure changes and yields quantitative data on domain/domain interactions. An experimental setup designed for attenuated total reflection and a specific approach for the analysis of the results is described. The study of one membrane protein, the gastric H + ,K + ‐ATPase, demonstrates the usefulness of 1 H/ 2 H exchange kinetics for the understanding of the molecular movement related to the catalytic activity. © 2004 Wiley Periodicals, Inc. Biopolymers, 2004