Purification of a capillary permeability increasing-enzyme from the venom of Agkistrodon caliginosus (kankoku-mamushi).

A capillary permeability increasing-enzyme was purified from the venom of Agkistrodon caliginosus by gel-filtration on Sephadex G-100, ion exchange chromatographies on CM-Sephadex C-50 and DEAE-Sephadex A-50 and affinity chromatography on p-aminobenzamidine-epsilon-aminocaproic acid-Sepharose 4B. By this procedure, 6.2 mg of the purified enzyme was obtained from 4 g of the venom. The purified enzyme was homogeneous by disc electrophoresis at pH 8.3 and pH 4.5, and did not show any caseinolytic, clotting or bradykinin-releasing activities. The enzyme hydrolyzed N-alpha-tosyl-L-arginine methylester and the specific activity of the enzyme was 7.3 N-alpha-tosyl-L-arginine methylester units per mg of protein. When 3 micrograms of the enzyme was injected into the depilated skin of the back of a rabbit, capillary permeability was increased.