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Isotope effects in peptide group hydrogen exchange
Author(s) -
Connelly Gregory P.,
Bai Yawen,
Jeng MeiFen,
Englander S. Walter
Publication year - 1993
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.340170111
Subject(s) - chemistry , reaction rate constant , solvent , kinetic isotope effect , equilibrium constant , hydrogen , peptide , isotope , amide , kinetics , stereochemistry , inorganic chemistry , deuterium , organic chemistry , atomic physics , biochemistry , physics , quantum mechanics
Kinetic and equilibrium isotope effects in peptide group hydrogen exchange reactions were evaluated. Unlike many other reactions, Kinetic isotope effects in amide hydrogen exchange are small because exchange pathways are not limited by bondbreaking steps. Rate constants for the acid‐cat‐alyzed exchange of peptide group NH, ND, and NT in H 2 O are essentially identical, but a solvent esotope effect doubles the rate in D 2 O. Rate constants for base‐catalyzed exchange in H 2 O decrease slowly in the order NH > ND > NT. The alkaline rate constant in D 2 O is very close to that in H 2 O when account is taken of the glass electrode pH artifact and the difference in solvent ionization constant. Small equilibrium isotope effects lead to an excess equilibrium accumulation of the heavier isotopes by the peptide group. Results obtained are expressed in terms of rate constants for the random coil polypeptide, poly‐DL‐alanine, to provide reference rates for protein hydrogen exchange studies as described in Bai et al. [preceding paper in this issue]. © 1993 Wiley‐Liss, Inc.