Purification and characterization of methylmalonyl-CoA mutase from a photosynthetic coccolithophorid alga, Pleurochrysis carterae.
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.
Miyamoto
Emi
E
Department of Health Science, Kochi Women's University, 5-15 Eikokuji-cho, Kochi 780-8515, Japan.
Watanabe
Fumio
F
Yamaguchi
Yuji
Y
Takenaka
Hiroyuki
H
Nakano
Yoshihisa
Y
D001671:Biochemistry / Q000379:methods; D003038:Cobamides / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D056890:Eukaryota / Q000201:enzymology* / Q000502:physiology; D008765:Methylmalonyl-CoA Mutase / Q000302:isolation & purification* / Q000378:metabolism*; D008970:Molecular Weight; D010788:Photosynthesis; D013696:Temperature
D003038:Cobamides; D008765:Methylmalonyl-CoA Mutase