title: Purification and characterization of two distinct Ca2+-activated proteinases from hearts of hypertensive rats.
abstract: Two distinct Ca2+-activated proteinases were purified and characterized from hearts of hypertensive rats. Ca2+-activated proteinases I and II, having low and high Ca2+ requirements, respectively, were first separated by DEAE-cellulose chromatography. The enzymes were then purified individually by different column procedures: chromatography on phenyl-Sepharose, then Sephadex G-200 for proteinase I and reactive-red agarose for proteinase II. The apparent molecular weight of purified proteinase I was 125 000 and that for purified proteinase II was 110 000. Both enzymes are heterodimers made up of a larger catalytic subunit and a smaller subunit devoid of proteinase activity. Ca2+ concentrations for half-maximal activation were 5 microM for proteinase I and 200 microM for proteinase II. Both enzymes were inhibited by sulfhydryl-modifying agents, but exhibited different characteristics in the auto-digestion reaction in the presence of Ca2+. Proteinases I and II were also purified from hearts of normotensive rats and shown to be identical to their respective counterparts from hearts of hypertensive rats. However, proteinase II activity in hypertensive rat hearts was significantly elevated as compared to controls.
authors.forename: M
authors.initials: M
authors.lastname: Spalla
authors.forename: W
authors.initials: W
authors.lastname: Tsang
authors.forename: T H
authors.initials: TH
authors.lastname: Kuo
authors.forename: F
authors.initials: F
authors.lastname: Giacomelli
authors.forename: J
authors.initials: J
authors.lastname: Wiener
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002154:Calpain; D002364:Caseins / Q000378:metabolism; D002845:Chromatography; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D006977:Hypertension, Renal / Q000201:enzymology*; D007700:Kinetics; D046911:Macromolecular Substances; D008297:Male; D008970:Molecular Weight; D009206:Myocardium / Q000201:enzymology*; D011480:Protease Inhibitors; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D002364:Caseins; D046911:Macromolecular Substances; D011480:Protease Inhibitors; C036307:acetylcasein; D010450:Endopeptidases; D002154:Calpain; D002118:Calcium