Purification and properties of five different forms of human procarboxypeptidases.
Three different procarboxypeptidases A and two different procarboxypeptidases B have been isolated for the first time, in a pure and native state, from human pancreatic extracts. These proteins were purified in one or two quick steps by anion-exchange HPLC. All these forms have been biochemically characterized. Two of the procarboxypeptidases A, the A1 and A2 forms, are obtained in a monomeric state while the other, the A3 form, is obtained as a binary complex of a procarboxypeptidase A with a proproteinase E. This complex is stable in aqueous buffers at various ionic strengths and develops carboxypeptidase A and proteinase E activities in the presence of trypsin. The A1 and A2 forms show clear differences in electrophoretic mobility in SDS/polyacrylamide gels, isoelectric point, proteolytic activation process with trypsin and susceptibility to thermal denaturation. In contrast, these properties are similar in the A1 and A3 (binary complex) forms. On the other hand, with respect to the properties listed above, the B1 and B2 forms differ from each other mainly in isoelectric point. An overall comparison of the above properties reveals the unusual character of the A2 form, midway between the other A and B forms. N-terminal extended sequence analysis carried out on these proenzymes confirm that they constitute different isologous forms.
Pascual
R
R
Departament de Bioquímica i Biologia Molecular, Facultat de Ciències, Universitat Autònoma de Barcelona, Spain.
Burgos
F J
FJ
Salva
M
M
Soriano
F
F
Mendez
E
E
Aviles
F X
FX
D000818:Animals; D001483:Base Sequence; D043424:Carboxypeptidase B; D002268:Carboxypeptidases / Q000145:classification / Q000302:isolation & purification*; D043422:Carboxypeptidases A; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D004792:Enzyme Precursors / Q000145:classification / Q000302:isolation & purification*; D006358:Hot Temperature; D006801:Humans; D007526:Isoelectric Point; D010179:Pancreas / Q000201:enzymology*; D014357:Trypsin / Q000494:pharmacology
D004792:Enzyme Precursors; D002268:Carboxypeptidases; D043422:Carboxypeptidases A; D043424:Carboxypeptidase B; D014357:Trypsin