Procarboxypeptidase in rat pancreas. Overall characterization and comparison of the activation processes.
Three monomeric procarboxypeptidases and a binary complex consisting of a procarboxypeptidase and a chymotrypsinogen have been isolated from rat pancreas by HPLC. N-terminal sequence determination, substrate-specificity analysis and physico-chemical characterization showed that the carboxypeptidase precursors were the A1, A2 and B forms. No isomorphism could be detected for any of these proenzymes and no clear evidence was obtained for the presence of procarboxypeptidase-containing quaternary complexes of the types previously described for other species. Instead, we observed the presence of a binary complex between procarboxypeptidase A2 and chymotrypsinogen B. Among the major pancreatic endoproteinases, only trypsin was found to be a general activator of rat procarboxypeptidases in vitro. Time-course analysis of the products generated after trypsin addition confirmed that full activation of procarboxypeptidase A1 requires several cleavages in the C-terminal region (residues 87-94) of the activation segment, while procarboxypeptidases A2 and B require a single cleavage each. The carboxypeptidases released participate in the trimming of the activation segment in A1 and B, but not in A2, probably because of the high specificity of the latter in the active form.
Oppezzo
O
O
Institut de Biologia Fonamental, Universitat Autònoma de Barcelona, Spain.
Ventura
S
S
Bergman
T
T
Vendrell
J
J
Jörnvall
H
H
Avilés
F X
FX
D000595:Amino Acid Sequence; D000818:Animals; D002268:Carboxypeptidases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000302:isolation & purification / Q000378:metabolism*; D006358:Hot Temperature; D006868:Hydrolysis; D007700:Kinetics; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology*; D011489:Protein Denaturation; D051381:Rats; D011919:Rats, Inbred Strains
D004792:Enzyme Precursors; D002268:Carboxypeptidases