Purification and properties of a kininogenin from the venom of Lachesis muta (bushmaster).

An acidic kininogenin from Lachesis muta snake venom was purified to apparent homogeneity by a combination of gel filtration, isoelectric focusing and preparative gel electrophoresis. It was shown to be a highly stable serine protease (mol. wt 27,900; pI 5.4) capable of releasing bradykinin from low mol. wt bovine kininogen and of cleaving some synthetic chromogenic peptides with the following catalytic efficiencies (Kcat/Km, M-1.sec-1): N-benzoyl-Phe-Val-Arg-p-nitroanilide (1.92 x 10(4)); H-D-Val-Leu-Arg-p-nitroanilide (1.55 x 10(4)); N-acetyl-Phe-Arg-p-nitroanilide (3.98 x 10(2)); no hydrolysis was observed with N-benzoyl-Arg-p-nitroanilide. A marked and sustained hypotensive effect was recorded following i.v. injection of purified kininogenin into rats. Tachyphylaxis was observed after repeated i.v. injection of the enzyme, a phenomenon accompanied by a decrease of only 15% in the total circulating rat kininogen. Both the in vivo action and the enzymatic properties of the L. muta kininogenin indicate that this enzyme might be helpful for understanding the kinin-kininogen system.