IcmF Is a Fusion Between the Radical B12 Enzyme isobutyryl-CoA Mutase and Its G-protein Chaperone

J Biol Chem. 2010 Jan 1;285(1):655-66. doi: 10.1074/jbc.M109.062182. Epub 2009 Oct 28.

Abstract

Coenzyme B(12) is used by two highly similar radical enzymes, which catalyze carbon skeleton rearrangements, methylmalonyl-CoA mutase and isobutyryl-CoA mutase (ICM). ICM catalyzes the reversible interconversion of isobutyryl-CoA and n-butyryl-CoA and exists as a heterotetramer. In this study, we have identified >70 bacterial proteins, which represent fusions between the subunits of ICM and a P-loop GTPase and are currently misannotated as methylmalonyl-CoA mutases. We designate this fusion protein as IcmF (isobutyryl-CoA mutase fused). All IcmFs are composed of the following three domains: the N-terminal 5'-deoxyadenosylcobalamin binding region that is homologous to the small subunit of ICM (IcmB), a middle P-loop GTPase domain, and a C-terminal part that is homologous to the large subunit of ICM (IcmA). The P-loop GTPase domain has very high sequence similarity to the Methylobacterium extorquens MeaB, which is a chaperone for methylmalonyl-CoA mutase. We have demonstrated that IcmF is an active ICM by cloning, expressing, and purifying the IcmFs from Geobacillus kaustophilus, Nocardia farcinica, and Burkholderia xenovorans. This finding expands the known distribution of ICM activity well beyond the genus Streptomyces, where it is involved in polyketides biosynthesis, and suggests a role for this enzyme in novel bacterial pathways for amino acid degradation, myxalamid biosynthesis, and acetyl-CoA assimilation.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacteria / enzymology*
  • Bacteria / genetics
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Cobamides / metabolism
  • Computational Biology
  • Electron Spin Resonance Spectroscopy
  • Enzyme Activation
  • GTP Phosphohydrolases / metabolism
  • GTP-Binding Proteins / chemistry
  • GTP-Binding Proteins / metabolism*
  • Gene Expression Regulation, Bacterial
  • Isomerases / genetics
  • Isomerases / metabolism*
  • Kinetics
  • Molecular Chaperones / metabolism*
  • Nucleotides / metabolism
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Streptomyces / enzymology
  • Vitamin B 12 / metabolism*

Substances

  • Bacterial Proteins
  • Cobamides
  • Molecular Chaperones
  • Nucleotides
  • Recombinant Fusion Proteins
  • GTP Phosphohydrolases
  • GTP-Binding Proteins
  • Isomerases
  • cobamamide
  • Vitamin B 12