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Quantitative ¹³C and ²H NMR Relaxation Studies of the 723-Residue Enzyme Malate Synthase G Reveal a Dynamic Binding Interface^†

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Vitali Tugarinov and Lewis E. Kay*

Departments of Medical Genetics, Biochemistry, and Chemistry, University of Toronto, Toronto, Ontario, Canada M5S 1A8

Biochemistry, 2005, 44 (49), pp 15970–15977

DOI: 10.1021/bi0519809

Publication Date (Web): November 16, 2005

†

This work was supported by a grant from the Canadian Institutes of Health Research (CIHR) to L.E.K. V.T. acknowledges the financial support of the CIHR in the form of a postdoctoral fellowship. L.E.K. holds a Canada Research Chair in Biochemistry.

To whom correspondence should be addressed. E-mail: kay@pound.med.utoronto.ca. Fax: (416) 978-6885. Telephone: (416) 978-0741.

Abstract

A detailed understanding of molecular recognition is predicated not only on high-resolution static structures of the free and bound states but also on information about how these structures change with time, that is, molecular dynamics. Here we present a deuterium (²H) and carbon (¹³C) NMR relaxation study of methyl side chain dynamics in the 82 kDa enzyme malate synthase G (MSG) that is a promising target for the development of new antibiotic agents. It is shown that excellent agreement between ²H- and ¹³C-derived measures of dynamics is obtained, with correlation coefficients exceeding 0.95. The binding interface formed by MSG and its substrates is found to be highly dynamic in the ligand-free state of the enzyme with rigidification upon binding substrate. This study establishes that detailed, quantitative information about methyl side chain dynamics can be obtained by NMR on proteins with molecular masses on the order of 100 kDa and opens up the possibilities for studies of motion in a large number of important systems.

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History

Published In Issue December 13, 2005
Received September 29, 2005
Revised Manuscript Received November 1, 2005

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Accelerated Publication

Quantitative ¹³C and ²H NMR Relaxation Studies of the 723-Residue Enzyme Malate Synthase G Reveal a Dynamic Binding Interface^†