Direct Brønsted analysis of the restoration of activity to a mutant enzyme by exogenous amines

Michael D. Toney, Jack F. Kirsch

Research output: Contribution to journalArticle

208 Citations (Scopus)

Abstract

A true Brønsted analysis of proton transfer in an enzyme mechanism is made possible by the chemical rescue of an inactive mutant of aspartate aminotransferase, where the endogenous general base, Lys258, is replaced with Ala by site-directed mutagenesis. Catalytic activity is restored to this inactive mutant by exogenous amines. The eleven amines studied generate a Brønsted correlation with β of 0.4 for the transamination of cysteine suilfinate, when steric effects are included in the regression analysis. Localized mutagenesis thus allows the classical Brønsted analysis of transition-state structure to be applied to enzyme-catalyzed reactions.

Original languageEnglish (US)
Pages (from-to)1485-1488
Number of pages4
JournalScience
Volume243
Issue number4897
StatePublished - 1989

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Amines
Enzymes
Aspartate Aminotransferases
Site-Directed Mutagenesis
Mutagenesis
Cysteine
Protons
Regression Analysis

ASJC Scopus subject areas

  • General

Cite this

Direct Brønsted analysis of the restoration of activity to a mutant enzyme by exogenous amines. / Toney, Michael D.; Kirsch, Jack F.

In: Science, Vol. 243, No. 4897, 1989, p. 1485-1488.

Research output: Contribution to journalArticle

Toney, MD & Kirsch, JF 1989, 'Direct Brønsted analysis of the restoration of activity to a mutant enzyme by exogenous amines', Science, vol. 243, no. 4897, pp. 1485-1488.
Toney, Michael D. ; Kirsch, Jack F. / Direct Brønsted analysis of the restoration of activity to a mutant enzyme by exogenous amines. In: Science. 1989 ; Vol. 243, No. 4897. pp. 1485-1488.
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