Purification and properties of succinyl-coenzyme A-3-oxo acid coenzyme A-transferase from sheep kidney.
CoA-transferase (succinyl-CoA-3-oxo acid CoA-transferase, EC 2.8.3.5) isolated from sheep kidney was purified to homogeneity. The purified enzyme has a specific activity of approx. 200 units/mg. A mol.wt. of 110000 was obtained by gel filtration on Sephadex G-200, and a lower mol.wt. of 102000 was determined by analytical ultracentrifugation. A sedimentation coefficient of 5.6S was also determined. A subunit mol.wt. of 56000 was obtained by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Isoelectric focusing of sheep kidney extracts indicated the presence of a single band of CoA-transferase activity with pI9.0. However, isoelectric focusing of purified CoA-transferase showed the presence of two peaks of CoA-transferase activity with pI values of 8.7 and 8.4, suggesting the presence of proteolytic activity during purification. Evidence for sheep kidney CoA-transferase being a dimer of two identical subunits has been obtained from sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, the amino acid composition, peptide 'mapping' and N-terminal analysis.
D000090:Acetoacetates; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D003065:Coenzyme A; D007525:Isoelectric Focusing; D007668:Kidney / Q000201:enzymology*; D008970:Molecular Weight; D012756:Sheep; D013386:Succinates; D013466:Sulfurtransferases / Q000302:isolation & purification
D000090:Acetoacetates; D000596:Amino Acids; D013386:Succinates; D013466:Sulfurtransferases; D003065:Coenzyme A