title: Use of lectin affinity chromatography for the purification of collagenase from human polymorphonuclear leukocytes.
abstract: Polymorphonuclear leukocytes (PMNLs) store collagenase in an inactive form in secretory granules. The enzyme can be activated in vitro by limited proteolysis or by sulfhydryl-modifying agents such as N-ethylmaleimide (NEM). We have enriched NEM-activated collagenase 820-fold using granule isolation, gel filtration, and wheat germ agglutinin (WGA)-agarose chromatography. The use of WGA-agarose resulted in a 55-fold enrichment of collagenase in a single step with very little loss of activity. The chromatographic behavior of collagenase on other lectin matrices was explored and gave information about the type of complex asparagine-linked oligosaccharide found on collagenase isolated from PMNLs.
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods; D004789:Enzyme Activation; D005033:Ethylmaleimide / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D003012:Microbial Collagenase / Q000097:blood* / Q000302:isolation & purification; D008970:Molecular Weight; D009504:Neutrophils / Q000201:enzymology*; D014909:Wheat Germ Agglutinins
chemical_list: D014909:Wheat Germ Agglutinins; D003012:Microbial Collagenase; D005033:Ethylmaleimide