title: Purification and partial characterization of a thiol proteinase activating prokallikrein from the rat kidney cortex.
abstract: The rat kidney cortex contains at least three kinds of prokallikrein-activating proteinase, and among these the one with the highest molecular weight was purified by a procedure including chromatography on CM-cellulose, concanavalin A-Sepharose, organomercurial-Sepharose 4B and Sephadex G-100. The resulting preparation was apparently homogeneous, as assessed by SDS-polyacrylamide gel electrophoresis. The molecular weight was estimated to be 57,000. The optimal pH for the activation of prokallikrein by the preparation, termed activator I, was around 4.5. Activator I was inhibited by E-64, iodoacetate and leupeptin, but not by PMSF and phosphoramidon. In immunodiffusion analysis, the antiserum to activator I formed an immunoprecipitin arc with the extract from the kidney cortex or submandibular gland, but not with that from the pancreas. These results indicate that activator I is a thiol proteinase with a molecular weight of 57,000. A proteinase immunologically identical with activator I appears to be present in the submandibular gland.
authors.affiliation: Department of Pharmacology, Osaka University of Pharmaceutical Sciences, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Nishii
authors.forename: M
authors.initials: M
authors.lastname: Takaoka
authors.forename: M
authors.initials: M
authors.lastname: Nakamura
authors.forename: Y
authors.initials: Y
authors.lastname: Takenobu
authors.forename: S
authors.initials: S
authors.lastname: Morimoto
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D003546:Cysteine Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism; D004789:Enzyme Activation; D005779:Immunodiffusion; D007672:Kidney Cortex / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D010179:Pancreas / Q000032:analysis; D011288:Prekallikrein / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013363:Submandibular Gland / Q000032:analysis
chemical_list: D000596:Amino Acids; D011288:Prekallikrein; D003546:Cysteine Endopeptidases