Acetyl CoA carboxylase: isolation and characterization of native biotin carboxyl carrier protein.
A large form of biotin carboxyl carrier protein (BCCP(L)) has been isolated from extracts of Escherichia coli. It has a minimal molecular weight of 20,000, according to its behavior on sodium dodecylsulfate-polyacrylamide gel electrophoresis, and contains approximately 1 mol of biotin per 22,000 g of protein. BCCP(L) exhibits K(m) values, in the biotin carboxylase and transcarboxylase half-reactions of acetyl CoA carboxylase, of 2 x 10(-7) M and 4 x 10(-7) M, respectively; these values are 50-100 times lower than those obtained with smaller forms of BCCP previously isolated. Electrophoresis of crude extracts of E. coli indicates that the major biotin-containing protein migrates at the same rate as BCCP(L), which suggests that BCCP(L) is the native form of BCCP in E. coli.
D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification*; D001710:Biotin / Q000032:analysis; D003065:Coenzyme A; D004589:Electrophoresis, Disc; D004926:Escherichia coli / Q000201:enzymology* / Q000254:growth & development; D007700:Kinetics; D008025:Ligases / Q000302:isolation & purification; D008970:Molecular Weight; D011485:Protein Binding; D014316:Tritium
D001426:Bacterial Proteins; D014316:Tritium; D001710:Biotin; D008025:Ligases; D003065:Coenzyme A