Purification and characterization of a methanol-induced cobamide-containing protein from Sporomusa ovata.
The major cobamide-containing protein from methanol-utilizing Sporomusa ovata was 8-fold enriched to apparent homogeneity. The protein exhibited a molecular mass of 40 kDa and of 38 kDa determined by gel filtration and by SDS-polyacrylamide gel electrophoresis, respectively. This finding indicates a monomeric protein structure. Monospecific polyclonal antisera raised against the protein did not cross react with another cobamide-containing protein from Sporomusa cells. Only the 40 kDa cobamide-containing protein was induced by methanol, since proteins from cells grown on 3,4-dimethoxybenzoate, betaine H2/CO2, or fructose showed faint or no cross reaction. Hence, the 40 kDa cobamide-containing protein is presumably involved in the methyl-transfer reaction of the methanol metabolism. The purified enzyme revealed 1.1 mol of p-cresolyl cobamide per mol of protein, but it lacked of iron-sulfur centers. Remarkably, the cofactor was firmly bound to its protein.
Stupperich
E
E
Abteilung Angewandte Mikrobiologie, Universität Ulm, Federal Republic of Germany.
Aulkemeyer
P
P
Eckerskorn
C
C
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D002850:Chromatography, Gel; D003038:Cobamides / Q000032:analysis; D003067:Coenzymes / Q000302:isolation & purification; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D006089:Gram-Negative Anaerobic Bacteria / Q000737:chemistry* / Q000378:metabolism; D000432:Methanol / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight
D001426:Bacterial Proteins; D003038:Cobamides; D003067:Coenzymes; D000432:Methanol