Human pancreatic carboxypeptidase B.  I.  Isolation, purification, and characterization of fraction II.
Human carboxypeptidase B fraction II has been purified from pancreatic juice by DEAE-'Sephadex' chromatography, isoelectric focusing, and 'Sephadex' G-100 gel filtration. The enzyme has been characterized by analytical polyacrylamide disc-gel electrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, amino acid analysis Km determination, molecular weight determination on 'Sephadex' G-100, zinc analysis, and inhibition by metal chelating agents. Human carboxypeptidase B fraction II appeared homogeneous in analytical polyacrylamide disc-gel electrophoresis, but showed two components of 23,500 and 9,200 daltons in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Zinc analysis revealed 0.96 gram atoms of zinc per mole of enzyme, and a Km of 65 +/- 3 muM was determined for hydrolysis of hippuryl-L-arginine.
D000596:Amino Acids / Q000032:analysis; D002268:Carboxypeptidases / Q000302:isolation & purification* / Q000378:metabolism; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004492:Edetic Acid / Q000494:pharmacology; D004589:Electrophoresis, Disc; D006801:Humans; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D010179:Pancreas / Q000201:enzymology*; D010618:Phenanthrolines / Q000494:pharmacology; D015032:Zinc / Q000032:analysis
D000596:Amino Acids; D010618:Phenanthrolines; D004492:Edetic Acid; D002268:Carboxypeptidases; D015032:Zinc