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Enzyme conformational dynamics during catalysis and in the ‘resting state’ monitored by hydrogen/deuterium exchange mass spectrometry
Author(s) -
Liu Yu-Hong,
Konermann Lars
Publication year - 2006
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2006.08.042
Subject(s) - hydrogen–deuterium exchange , chemistry , kinetics , deuterium , substrate (aquarium) , mass spectrometry , enzyme , enzyme kinetics , enzyme catalysis , carboxypeptidase a , steady state (chemistry) , amide , carboxypeptidase , chromatography , active site , biochemistry , physics , oceanography , quantum mechanics , geology
This work reports the use of electrospray mass spectrometry for studying the conformational dynamics of enzymes by amide hydrogen/deuterium exchange (HDX) measurements. A rapid‐mixing quench‐flow approach allows comparisons to be made between the HDX kinetics of free enzymes with those under steady‐state conditions. Experiments carried out on carboxypeptidase B in the absence of substrate and in the presence of saturating concentrations of hippuryl‐Arg result in HDX kinetics that are indistinguishable. This finding implies that the conformational dynamics that mediate HDX are not significantly different in the resting state of the enzyme and during substrate turnover.