Structure-based mutagenesis of the malonyl-CoA

acyl carrier protein transacylase from Streptomyces coelicolor

Andrew T Koppisch, Chaitan Khosla

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Malonyl-CoA:acyl carrier protein transacylase (MAT) provides acyl-ACP thioesters for the biosynthesis of aromatic polyketides, and thus is the primary gatekeeper of substrate specificity in type II PKS. A recent report described the X-ray crystal structure of the Streptomyces coelicolor MAT and suggested active site residues which may be important for substrate selectivity [Keatinge-Clay, A. T., et al. (2003) Structure 11, 147-154]. Mutants were made to test the proposed roles of these residues, and the enzymes were characterized kinetically with respect to native and non-native substrates. The activity of the MAT was observed to be greatly attenuated in many of the observed mutants; however, the Km for malonyl-CoA was only modestly affected. Our results suggest the MAT uses an active site that is rigorously ordered around the acyl-thioester moiety of the acyl-CoA to facilitate rapid and efficient transacylation to an ACP. Our results also suggest that the MAT does not discriminate against α-substituted acyl-CoA thioesters solely on the basis of substrate size.

Original languageEnglish (US)
Pages (from-to)11057-11064
Number of pages8
JournalBiochemistry
Volume42
Issue number37
DOIs
StatePublished - Sep 23 2003
Externally publishedYes

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Acyl Carrier Protein
Malonyl Coenzyme A
Streptomyces coelicolor
Acyl Coenzyme A
Mutagenesis
Catalytic Domain
Polyketides
Substrates
Substrate Specificity
X-Rays
Biosynthesis
Enzymes
Crystal structure
X rays
clay

ASJC Scopus subject areas

  • Biochemistry

Cite this

Structure-based mutagenesis of the malonyl-CoA : acyl carrier protein transacylase from Streptomyces coelicolor. / Koppisch, Andrew T; Khosla, Chaitan.

In: Biochemistry, Vol. 42, No. 37, 23.09.2003, p. 11057-11064.

Research output: Contribution to journalArticle

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