Quaternary structure of the oxaloacetate decarboxylase membrane complex and mechanistic relationships to pyruvate carboxylases

J Biol Chem. 2011 Mar 18;286(11):9457-67. doi: 10.1074/jbc.M110.197442. Epub 2011 Jan 5.

Abstract

The oxaloacetate decarboxylase primary Na(+) pump (OAD) is an essential membrane protein complex that functions in the citrate fermentation pathway of some pathogenic bacteria under anaerobic conditions. OAD contains three different subunits: Oad-α, a biotinylated extrinsic protein that catalyzes the α-ketodecarboxylation of oxaloacetate; Oad-γ, a structural bitopic membrane protein whose cytosolic tail (named as Oad-γ') binds tightly to Oad-α; and Oad-β, a multispan transmembrane α-helical protein that constitutes the Na(+) channel. How OAD is organized structurally at the membrane and what the molecular determinants are that lead to an efficient energy coupling mechanism remain elusive. In the present work, we elucidate the stoichiometry of the native complex as well as the low resolution structure of the peripheral components of OAD (Oad-α and Oad-γ') by small angle x-ray scattering. Our results point to a quaternary assembly similar to the pyruvate carboxylase complex organization. Herein, we propose a model in which the association in pairs of Oad-α dimers, mediated by Oad-γ, results in the acquisition of a functional oligomeric state at the bacterial membrane. New structural insights for the conformational rearrangements associated with the carboxylbiotin transfer reaction within OAD are provided.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Carboxy-Lyases / chemistry*
  • Carboxy-Lyases / genetics
  • Membrane Proteins / chemistry*
  • Membrane Proteins / genetics
  • Protein Structure, Quaternary
  • Protein Structure, Secondary
  • Pyruvate Carboxylase / chemistry*
  • Pyruvate Carboxylase / genetics
  • Vibrio cholerae / enzymology*
  • Vibrio cholerae / genetics

Substances

  • Bacterial Proteins
  • Membrane Proteins
  • Carboxy-Lyases
  • oxaloacetate decarboxylase
  • Pyruvate Carboxylase