Purification and characterization of methylmalonyl-CoA mutase from a photosynthetic coccolithophorid alga, Pleurochrysis carterae

Comp Biochem Physiol B Biochem Mol Biol. 2004 Jun;138(2):163-7. doi: 10.1016/j.cbpc.2004.03.006.

Abstract

Low activity (about 4 mU/mg protein) of 5'-deoxyadenosylcobalamin-dependent methylmalonyl-CoA mutase (MCM; EC 5.4.99.2) was found in a cell homogenate of a photosynthetic coccolithophorid alga, Pleurochrysis carterae. Most of the enzyme occurred as the apo-enzyme, which was labile during purification. The holo-enzyme, which was converted from the apo-enzyme by incubation with 10 microM 5'-deoxyadenosylcobalamin at 4 degrees C in the dark, was purified to homogeneity and partially characterized. An apparent molecular mass for the enzyme of 150+/-5 kDa was calculated by Superdex 200 pg gel filtration. SDS-polyacrylamide gel electrophoresis of the purified enzyme gave a single protein band with an apparent molecular mass of 80+/-5 kDa, indicating that the P. carterae enzyme occurs as a homodimer. Some properties of methylmalonyl-CoA mutase from P. carterae were studied.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biochemistry / methods
  • Cobamides / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Eukaryota / enzymology*
  • Eukaryota / physiology
  • Methylmalonyl-CoA Mutase / isolation & purification*
  • Methylmalonyl-CoA Mutase / metabolism*
  • Molecular Weight
  • Photosynthesis
  • Temperature

Substances

  • Cobamides
  • Methylmalonyl-CoA Mutase