title: Purification and characterization of a 27,000-Mr extracellular proteinase from Trichophyton rubrum.
abstract: A proteinase of Mr 27,000 with a possible role in the metabolism and invasion of host tissues was purified from the conditioned medium of Trichophyton rubrum by concanavalin A and anion-exchange chromatography. Peaks of proteolytic activity were analyzed by substrate gel electrophoresis. The 27,000-Mr proteinase had a pH optimum of 8.0, a calcium dependence of 2 mM, and was inhibited by serine proteinase inhibitors, especially phenylmethylsulfonyl fluoride and Phe-Gly-Ala-Leu-chloromethyl ketone. By polyacrylamide gel electrophoresis, the 27,000-Mr proteinase had a reduced molecular weight of 44,000 and reacted with [3H]diisopropyl fluorophosphate. The proteinase degraded azocoll, type III collagen, type IV procollagen, laminin, fibronectin, and the peptide substrates succinyl-Ala-Ala-Pro-Phe-p-nitroanilide (1,573 M-1 s-1) and t-butyloxy carbonyl-Ala-Ala-Leu-p-nitroanilide (1,614 M-1 s-1).
authors.affiliation: Department of Pathology, University of California, San Francisco, 94143.
authors.forename: G
authors.initials: G
authors.lastname: Apodaca
authors.forename: J H
authors.initials: JH
authors.lastname: McKerrow
mesh_terms: D002118:Calcium / Q000502:physiology; D010450:Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism / Q000502:physiology; D005110:Extracellular Space / Q000201:enzymology* / Q000382:microbiology; D005656:Fungal Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000502:physiology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D046911:Macromolecular Substances; D008970:Molecular Weight; D009842:Oligopeptides / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity; D014249:Trichophyton / Q000201:enzymology* / Q000502:physiology
chemical_list: D005656:Fungal Proteins; D046911:Macromolecular Substances; D009842:Oligopeptides; D011480:Protease Inhibitors; D010450:Endopeptidases; D002118:Calcium