Biochemical and structural studies of malate synthase from Mycobacterium tuberculosis

Clare V. Smith, Chih Chin Huang, Andras Miczak, David G. Russell, James C. Sacchettini, Kerstin Höner zu Bentrup

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Establishment or maintenance of a persistent infection by Mycobacterium tuberculosis requires the glyoxylate pathway. This is a bypass of the tricarboxylic acid cycle in which isocitrate lyase and malate synthase (GlcB) catalyze the net incorporation of carbon during growth of microorganisms on acetate or fatty acids as the primary carbon source. The glcB gene from M. tuberculosis, which encodes malate synthase, was cloned, and GlcB was expressed in Escherichia coli. The influence of media conditions on expression in M. tuberculosis indicated that this enzyme is regulated differentially to isocitrate lyase. Purified GlcB had Km values of 57 and 30 μM for its substrates glyoxylate and acetyl coenzyme A, respectively, and was inhibited by bromopyruvate, oxalate, and phosphoenolpyruvate. The GlcB structure was solved to 2.1-Å resolution in the presence of glyoxylate and magnesium. We also report the structure of GlcB in complex with the products of the reaction, coenzyme A and malate, solved to 2.7-Å resolution. Coenzyme A binds in a bent conformation, and the details of its interactions are described, together with implications on the enzyme mechanism.

Original languageEnglish
Pages (from-to)1735-1743
Number of pages9
JournalJournal of Biological Chemistry
Issue number3
Publication statusPublished - Jan 17 2003


ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Smith, C. V., Huang, C. C., Miczak, A., Russell, D. G., Sacchettini, J. C., & Höner zu Bentrup, K. (2003). Biochemical and structural studies of malate synthase from Mycobacterium tuberculosis. Journal of Biological Chemistry, 278(3), 1735-1743.