Preparative isolation of the two forms of pig pancreatic pro-(carboxypeptidase A) and their monomeric carboxypeptidases A.
A method is reported for the preparative isolation of the two forms of pro-(carboxypeptidase A) from pig pancreas: the monomer and the binary complex with pro-(proteinase E). This method, which is mainly based on chromatography on DEAE-Sepharose at pH 5.7, allows these proenzymes to be prepared more quickly and in safer conditions than with other reported methods. Undegraded and homogeneous carboxypeptidase A1 and A2 species (peptidyl-L-amino acid hydrolase, EC 3.4.17.1), in monomeric forms with high specific activity, are also obtained in high yield by controlled trypsin activation of either of the pro-(carboxypeptidases A) followed by chromatography on DEAE-Sepharose at pH 5.8 under dissociating conditions (7 M-urea).
D000818:Animals; D002268:Carboxypeptidases / Q000302:isolation & purification*; D043422:Carboxypeptidases A; D002846:Chromatography, Affinity; D004792:Enzyme Precursors / Q000302:isolation & purification*; D007527:Isoenzymes / Q000302:isolation & purification*; D046911:Macromolecular Substances; D008722:Methods; D010179:Pancreas / Q000201:enzymology; D013552:Swine; D014508:Urea
D004792:Enzyme Precursors; D007527:Isoenzymes; D046911:Macromolecular Substances; D014508:Urea; D002268:Carboxypeptidases; D043422:Carboxypeptidases A