Studies on the prekallikrein (kallikreinogen)--kallikrein enzyme system of human plasma. I. Isolation and purification of plasma kallikreins.
By measurement of its arginine esterase activity, plasma kallikrein was purified from fresh frozen ACD plasma. The steps involved alcohol fractionation, isoelectric precipitation, and carboxymethyl (CM) Sephadex and DEAE cellulose chromatography. Three enzymatically active fractions were finally isolated and termed plasma kallikreins I, II, and III; they represented purifications of 970,320- and 590-fold, respectively. All three kallikreins were active biologically; they increased vascular permeability in the guinea pig and released a kinin from human plasma, as measured in the rat uterus bioassay. Bradykinin and/or closely related kinins were identified in the kallikrein I plasma digest by radioimmunoassay. Kallikreins I, II, and III had similar ratios of hydrolytic activity on a variety of arginine and lysine esters and were immunochemically related. However, differences were present on physicochemical characterization: kallikrein I had S(20,[unk]) of 5.7, a mol wt of 99,800, and migrated as a slow gamma globulin; kallikrein II migrated as a fast gamma globulin with a mol wt of 163,000, but the evidence suggested that it was closely related, if not interconvertible, with kallikrein I. Kallikrein III, on the other hand, migrated as an alpha globulin and reacted quite differently with inhibitors.
D000818:Animals; D001681:Biological Assay; D002199:Capillary Permeability / Q000187:drug effects; D002482:Cellulose; D011232:Chemical Precipitation; D002845:Chromatography; D002850:Chromatography, Gel; D004586:Electrophoresis; D004792:Enzyme Precursors / Q000302:isolation & purification*; D004950:Esterases / Q000032:analysis; D005260:Female; D006168:Guinea Pigs; D006801:Humans; D005779:Immunodiffusion; D007610:Kallikreins / Q000302:isolation & purification*; D008970:Molecular Weight; D011863:Radioimmunoassay; D014461:Ultracentrifugation; D014599:Uterus / Q000187:drug effects
D004792:Enzyme Precursors; D002482:Cellulose; D004950:Esterases; D007610:Kallikreins