Preparation and properties of ornithine-oxo-acid aminotransferase of rat kidney. Comparison with the liver enzyme.
Ornithine-oxo-acid aminotransferase (EC 2.6.1.13) from rat kidney was prepared as a single homogeneous protein as judged by polyacrylamide gel electrophoresis, ultracentrifuge analysis and double diffusion precipitin test. Content of pyridoxal phosphate, light absorption spectra, circular dicroism spectra, Km values, inhibitors, and electrophoretic mobilities of the proteins after reactions with group modifying reagents were similar for the ornithine-oxo-acid aminotransferases of rat kidney and liver. Rates of reaction with group modifying reagents, stabilities to storage at -15 degrees C, and stabilities to temperatures above 55 degrees C differed significantly for the two enzymes. The liver enzyme contained two more cysteine residues than the kidney enzyme as determined by three different methods. Heating the liver enzyme at 66-67 degrees C at pH 5.9 for 1 h decreased the thiol groups titratable by 5,5'-dithio-bis(2-nitrobenzoic acid) (Nbs2). Uncer the same conditions titratable thiol groups of the kidney enzyme were not decreased. Amino acid analysis revealed probably significant differences in tyrosine and isoleucine content in addition to cysteine. It was concluded that the primary structures of ornithine-oxo-acid aminotransferases of rat liver and kidney are not fully identical.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002172:Canavanine / Q000494:pharmacology; D003545:Cysteine / Q000032:analysis; D007656:Ketoglutaric Acids / Q000494:pharmacology; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D009952:Ornithine / Q000494:pharmacology; D009953:Ornithine-Oxo-Acid Transaminase / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D011732:Pyridoxal Phosphate / Q000032:analysis; D051381:Rats; D013438:Sulfhydryl Compounds / Q000032:analysis; D013696:Temperature; D000637:Transaminases / Q000378:metabolism*
D000596:Amino Acids; D007656:Ketoglutaric Acids; D013438:Sulfhydryl Compounds; D002172:Canavanine; D011732:Pyridoxal Phosphate; D009952:Ornithine; D000637:Transaminases; D009953:Ornithine-Oxo-Acid Transaminase; D003545:Cysteine