title: Human carbamylphosphate synthetase I. Stabilization, purification, and partial characterization of the enzyme from human liver.
abstract: Carbamylphosphate synthetase I from human liver was stabilized, purified, and partially characterized. The labile enzyme was stabilized in cell-free extracts by the presence of MgATP and dithiothreitol at pH 7.8. The stabilized enzyme was purified by a rapid procedure consisting of ion exchange chromatograhy and electrofocusing The native molecular weight of the enzyme was determined by gel filtration to be 190,000. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated a monomeric molecular weight of 165,000. The isoelectric point of the purified enzyme was 6.05, and only one species of active enzyme was observed during electrofocusing of both purified enzyme preparations and crude liver homogenates. The enzyme exhibited a pH optimum of 7.8. The apparent Michaelis constants for NH4+, HCO3-, MgATP, and the activator, N-acetyl-L-glutamic acid, were 0.8, 6.7, 1.1, and 0.1 mM, respectively.
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000328:Adult; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000302:isolation & purification*; D004229:Dithiothreitol / Q000494:pharmacology; D004355:Drug Stability; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D010770:Phosphotransferases / Q000302:isolation & purification*
chemical_list: D000255:Adenosine Triphosphate; D010770:Phosphotransferases; D002222:Carbamoyl-Phosphate Synthase (Ammonia); D008274:Magnesium; D004229:Dithiothreitol