Purification and preliminary characterization of stratum corneum chymotryptic enzyme: a proteinase that may be involved in desquamation.
In recent work we have shown that a serine proteinase, stratum corneum chymotryptic enzyme, with properties compatible with a role in desquamation in vitro as well as in vivo, is generally present in human stratum corneum. The enzymologic properties of the stratum corneum chymotryptic enzyme in a KCl extract of dissociated plantar corneocytes were compared with those of other known chymotryptic serine proteinases. Stratum corneum chymotryptic enzyme was found to differ significantly from bovine chymotrypsin, human cathepsin G, and human mast cell chymases in regard to inhibitor profile and substrate specificity. Stratum corneum chymotryptic enzyme was further purified from KCl extracts of dissociated plantar corneocytes by affinity chromatography on gels with covalently linked soybean trypsin inhibitor. The purified preparation contained one major component with apparent molecular weight 25 kD and one minor component with slightly higher apparent molecular weight as revealed by Coomassie staining after electrophoresis in polyacrylamide gels with sodium dodecyl sulphate of samples that had not been reduced. Both these components were associated with chymotrypsin-like activity as revealed by zymography in polyacrylamide gels with co-polymerized casein. On zymography gels, the purified preparation was also found to contain minor amounts of components with trypsin-like activity. The major purified protein had an apparent molecular weight of around 28 kD after reduction and full denaturation and was shown to contain carbohydrate.
Egelrud
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Department of Dermatology, University Hospital, Umeå, Sweden.
D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002417:Cattle; D002846:Chromatography, Affinity; D006801:Humans; D007610:Kallikreins; D007641:Keratolytic Agents / Q000737:chemistry / Q000302:isolation & purification*; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000378:metabolism; D011489:Protein Denaturation; D012697:Serine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification*
D002241:Carbohydrates; D007641:Keratolytic Agents; D010504:Periodic Acid; C009288:metaperiodate; C082472:KLK7 protein, human; D007610:Kallikreins; D012697:Serine Endopeptidases