A Symmetrical Tetramer for S. aureus Pyruvate Carboxylase in Complex with Coenzyme A

Linda P.C. Yu, Song Xiang, Gorka Lasso, David Gil, Mikel Valle, Liang Tong

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Pyruvate carboxylase (PC) is a conserved metabolic enzyme with important cellular functions. We report crystallographic and cryo-electron microscopy (EM) studies of Staphylococcus aureus PC (SaPC) in complex with acetyl-CoA, an allosteric activator, and mutagenesis, biochemical, and structural studies of the biotin binding site of its carboxyltransferase (CT) domain. The disease-causing A610T mutation abolishes catalytic activity by blocking biotin binding to the CT active site, and Thr908 might play a catalytic role in the CT reaction. The crystal structure of SaPC in complex with CoA reveals a symmetrical tetramer, with one CoA molecule bound to each monomer, and cryo-EM studies confirm the symmetrical nature of the tetramer. These observations are in sharp contrast to the highly asymmetrical tetramer of Rhizobium etli PC in complex with ethyl-CoA. Our structural information suggests that acetyl-CoA promotes a conformation for the dimer of the biotin carboxylase domain of PC that might be catalytically more competent.

Original languageEnglish (US)
Pages (from-to)823-832
Number of pages10
JournalStructure
Volume17
Issue number6
DOIs
StatePublished - Jun 10 2009
Externally publishedYes

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Pyruvate Carboxylase
Carboxyl and Carbamoyl Transferases
Coenzyme A
Cryoelectron Microscopy
Acetyl Coenzyme A
biotin carboxylase
Biotin
Staphylococcus aureus
Rhizobium etli
Mutagenesis
Catalytic Domain
Binding Sites
Mutation
Enzymes

Keywords

  • PROTEINS

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

A Symmetrical Tetramer for S. aureus Pyruvate Carboxylase in Complex with Coenzyme A. / Yu, Linda P.C.; Xiang, Song; Lasso, Gorka; Gil, David; Valle, Mikel; Tong, Liang.

In: Structure, Vol. 17, No. 6, 10.06.2009, p. 823-832.

Research output: Contribution to journalArticle

Yu, Linda P.C. ; Xiang, Song ; Lasso, Gorka ; Gil, David ; Valle, Mikel ; Tong, Liang. / A Symmetrical Tetramer for S. aureus Pyruvate Carboxylase in Complex with Coenzyme A. In: Structure. 2009 ; Vol. 17, No. 6. pp. 823-832.
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