Specific cleavage of reduced and S-carboxamidomethylated neurophysin II by the collagenase of Clostridium histolyticum.
Purified collagenase of Clostridium histolyticum was shown to cleave reduced and S-carboxamidomethylated bovine neurophysin between Cys-13 and Gly-14. The scission resulted in formation of two separable fragments: a smaller peptide arising from residues 1 through 13, and a larger peptide comprising the remainder of the residues of the protein. By dansylation procedures, the smaller peptide was shown to have amino-terminal alanine as expected from the sequence of neurophysin II, and the larger peptide had amino-terminal glycine as anticipated. These results show that collagenase indeed cleaves bovine neurophysin II in accord with the specificity postulated for that enzyme, i.e., scission between -X-Gly- in a sequence of -Pro-X-Gly-Pro-Y-. This result, obtained with a non-collagenous protein substrate, is further confirmation of the specificity of collagenase as established by its action on collagens and on synthetic oligopeptides.
D000596:Amino Acids / Q000032:analysis; D003013:Clostridium / Q000201:enzymology*; D007460:Iodoacetamide; D003012:Microbial Collagenase / Q000302:isolation & purification; D009481:Neurophysins; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000032:analysis
D000596:Amino Acids; D009481:Neurophysins; D010446:Peptide Fragments; D003012:Microbial Collagenase; D007460:Iodoacetamide