Fibrinolytic protein from the venom of Calloselasma rhodostoma

Abstract

Malayan pit viper (Calloselasma rhodostoma) envenomation is a major health problem in south east Asian countries. Durign envenomation, the venom component mainly effects the hemostatic system and the venom also exhibits strong fibrinolytic activity. In this research, fibrinolytic enzyme from C. rhodostoma venom was purified in three step including hydrophobic interaction chromatography (HIC), size exclusion chromatography with protein-pak[superscript TM] 125 column and finally with protein-pak[superscript TM] 60 column. Electrophoresis titration curve of protein was investigated. The biological activities were determined including fibrinolytic and hemorrhagic, disintegrin and gelatinase activities. Purified fibrinolytic enzyme showed gelatinase and insulin b chain digestion without hemorrhagic and disintegrin activities. In additionally, insulin b chain digestion was inhibited by Virginia opossum (Didelphis verginianan) serum. This enzyme was characterized to be a fibrinogenase because it digest a chain of human fibrinogen. From Mass spectrometry analysis, This enzyme was composed of 236 amino acid residue with molecular weigh of 26701.5 Da. Three dimensional model revealing active site and post translation modification were calculated using swiss model program.