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Exploring steric constraints on protein mutations using MAGE/PROBE

Published online by Cambridge University Press:  15 December 2000

J. MICHAEL WORD
Affiliation:
Department of Biochemistry, Duke University, Durham, North Carolina 27710-3711 Present address: Glaxo Wellcome, Inc., Research Triangle Park, North Carolina 27709.
ROBERT C. BATEMAN
Affiliation:
Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, Mississippi 39406
BRENT K. PRESLEY
Affiliation:
Department of Biochemistry, Duke University, Durham, North Carolina 27710-3711 Present address: University of Texas Southwestern Medical School, Dallas, Texas 75219.
SIMON C. LOVELL
Affiliation:
Department of Biochemistry, Duke University, Durham, North Carolina 27710-3711
DAVID C. RICHARDSON
Affiliation:
Department of Biochemistry, Duke University, Durham, North Carolina 27710-3711
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Abstract

When planning a mutation to test some hypothesis, one crucial question is whether the new side chain is compatible with the existing structure; only if it is compatible can the interpretation of mutational results be straightforward. This paper presents a simple way of using the sensitive geometry of all-atom contacts (including hydrogens) to answer that question. The interactive MAGE/PROBE system lets the biologist explore conformational space for the mutant side chain, with an interactively updated kinemage display of its all-atom contacts to the original structure. The Autobondrot function in PROBE systematically explores that same conformational space, outputting contact scores at each point, which are then contoured and displayed. These procedures are applied here in two types of test cases, with known mutant structures. In ricin A chain, the ability of a neighboring glutamate to rescue activity of an active-site mutant is modeled successfully. In T4 lysozyme, six mutations to Leu are analyzed within the wild-type background structure, and their Autobondrot score maps correctly predict whether or not their surroundings must shift significantly in the actual mutant structures; interactive examination of contacts for the conformations involved explains which clashes are relieved by the motions. These programs are easy to use, are available free for UNIX or Microsoft Windows operating systems, and should be of significant help in choosing good mutation experiments or in understanding puzzling results.

Type
Research Article
Copyright
2000 The Protein Society

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