InforLorV4, Main, Exploration, bibRecord, 004A81

The mechanism of rate-limiting motions in enzyme function

Identifieur interne : 004A81 ( Main/Exploration ); précédent : 004A80; suivant : 004A82

The mechanism of rate-limiting motions in enzyme function

Auteurs : Eric D. Watt [États-Unis] ; Hiroko Shimada [États-Unis] ; Evgenii L. Kovrigin [États-Unis] ; J. Patrick Loria [États-Unis]

Source :

Proceedings of the National Academy of Sciences of the United States of America [ 0027-8424 ] ; 2007.

RBID : PMC:1924554

Abstract

The ability to use conformational flexibility is a hallmark of enzyme function. Here we show that protein motions and catalytic activity in a RNase are coupled and display identical solvent isotope effects. Solution NMR relaxation experiments identify a cluster of residues, some distant from the active site, that are integral to this motion. These studies implicate a single residue, histidine-48, as the key modulator in coupling protein motion with enzyme function. Mutation of H48 to alanine results in loss of protein motion in the isotope-sensitive region of the enzyme. In addition, k_cat decreases for this mutant and the kinetic solvent isotope effect on k_cat, which was 2.0 in WT, is near unity in H48A. Despite being located 18 Å from the enzyme active site, H48 is essential in coordinating the motions involved in the rate-limiting enzymatic step. These studies have identified, of ≈160 potential exchangeable protons, a single site that is integral in the rate-limiting step in RNase A enzyme function.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1924554

DOI: 10.1073/pnas.0702551104
PubMed: 17615241
PubMed Central: 1924554

Affiliations:

Le document en format XML

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The mechanism of rate-limiting motions in enzyme function

The mechanism of rate-limiting motions in enzyme function

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