Studies on phospho lipase a ec 188.8.131.52 in trimeresurus flavoviridis venom part 3 purification and some properties of phospho lipase a inhibitor in habu serum
Journal of Biochemistry (Tokyo) 80(2): 341-349
Phospholipase A [EC 184.108.40.206] inhibitor was purified from Habu (T. flavoviridis) serum by gel filtration on Sephadex G-200, chromatography on DE-23 cellulose and affinity chromatography on a Sepharose 4B-phospholipase A column. By these procedures, a 31-fold increase in specific activity was attained with a yield of 15%. The purified material was homogeneous as judged by cellulose acetate and polyacrylamide gel electrophoresis. It had an apparent MW of 100,000 as measured by gel filtration on Sephadex G-200. The purified inhibitor was stable for 20 min at 80.degree. and was unstable below pH 6. It migrated before albumin in cellulose acetate electrophoresis and did not form any precipitin line with the crude venom or with purified phospholipase A in immunodiffusion tests. An 8-fold excess of the purified inhibitor by weight was required to inhibit completely both the egg yolk clearing action and the hemolytic action of phospholipase A.