Reactions of alternate substrates demonstrate stereoelectronic control of reactivity in dialkylglycine decarboxylase

Shaoxian Sun, Roger F. Zabinski, Michael D. Toney

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29 Scopus citations

Abstract

Kinetic and product analyses of the reactions of dialkylglycine decarboxylase with several alternative substrates are presented. Rate constants for the reactions of amino and keto acids of several substrates decrease logarithmically with increasing side-chain size. Conversely, k(cat) for L-amino acid decarboxylation increases with side-chain size. These and other data confirm a proposed model for three binding subsites in the active site. In this model, bond making and breaking in both, tho decarboxylation and transamination half-reactions occurs at the 'A' subsite, which maintains the scissile bond aligned with the p orbitals of the conjugated aldimine and thus maximizes stereoelectronic effects. This strongly supports the proposal by Dunathan (Proc. Natl. Acad. Sci. U.S.A. 55, 712-716) that PLP-dependent enzymes can largely control reaction specificity by specific orientation about C(α) in the external aldimine intermediate. The 'B' subsite can accept either an alkyl or a carboxylate group, while the 'C' subsite accepts only small alkyl groups. This model predicts the existence of nonproductive binding modes for amino acids, which is proposed to be the ultimate origin of the k(cat) increase with side-chain size for L-amino acid decarboxylation. The specificity of the 2-aminoisobutyrate decarboxylation half-reaction toward oxidative decarboxylation is very high (< 1 in 105 turnovers yields nonoxidative decarboxylation). The origin of this specificity is explored with the reactions of amino- and methylaminomalonate. These substrates exhibit high yields of nonoxidative decarboxylation, providing support for a model in which the interaction between a carboxylate group in the B subsite and Arg406 is a prerequisite to proton donation to and removal from C(α).

Original languageEnglish (US)
Pages (from-to)3865-3875
Number of pages11
JournalBiochemistry
Volume37
Issue number11
DOIs
StatePublished - Mar 17 1998
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry

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