Application of affinity chromatography to the purification of collagenase for the isolation of insoluble elastin.
ClostrIdium histolyticum collagenase (clostridiopeptidase A, EC 3.4.4.19) was purified by batchwise separation with DEAE-cellulose followed by affinity chromatography on a column of alkali-treated elastin. The N-terminal amino acid profile of elastin isolated from bovine ligamentum nuchae using this enzyme preparation was compared with that of a duplicate sample purified with a mixture of collagenase I and II (Yoshida, E, and Noda, H. (1965) Biochim. Biopsys. Acta 105, 562-574). An approx. three-fold decrease in the molar concentration of N-terminal residues and a considerable reduction in their number was obtained by using the former enzyme preparation.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D003013:Clostridium / Q000201:enzymology; D004549:Elastin / Q000302:isolation & purification*; D006595:Hexosamines / Q000032:analysis; D006909:Hydroxyproline / Q000032:analysis; D008022:Ligaments / Q000032:analysis; D003012:Microbial Collagenase / Q000302:isolation & purification*; D012995:Solubility
D000596:Amino Acids; D006595:Hexosamines; D004549:Elastin; D003012:Microbial Collagenase; D006909:Hydroxyproline