Rabbit prekallikrein. Purification, biochemical characterization, and mechanism of activation.
Rabbit prekallikrein (RPK) was purified from rabbit plasma by ion exchange and lectin column chromatography and preparative polyacrylamide gel electrophoresis. A 1500-fold purification was routinely achieved with a final yield of 5-10%. The purified RPK was found to be a glycoprotein with an apparent molecular weight of 88,000. Activation of RPK with either trypsin or rabbit Hageman factor (active) occurs by limited proteolytic cleavage, producing two disulfide-linked polypeptide chains with molecular weights of 55,000 and 35,000. Both chains contain carbohyrate and the 35,000-molecular-weight polypeptide was shown to incorporate [3H]DFP. Activation of RPK in kaolin-treated plasma was shown to proceed by an analogous mechanism yielding 55,000- and 35,000-molecular-weight polypeptide chains.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002852:Chromatography, Ion Exchange / Q000379:methods; D003208:Concanavalin A; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004789:Enzyme Activation; D005174:Factor XII / Q000494:pharmacology; D007106:Immune Sera; D007610:Kallikreins / Q000302:isolation & purification*; D011288:Prekallikrein / Q000302:isolation & purification*; D011817:Rabbits; D014357:Trypsin / Q000494:pharmacology
D000596:Amino Acids; D007106:Immune Sera; D003208:Concanavalin A; D005174:Factor XII; D011288:Prekallikrein; D007610:Kallikreins; D014357:Trypsin