Protein compositions and biological activities of king cobra (Ophiophagus hannah) venom

Abstract

In this research involves comparison of protein composition and biological activities of different age and region of originate of three Ophiophagus hannah venoms. Biological activity was measured via cytotoxicity in cell line. KV2, crude venom showed higher toxicity than KV6 and KV9 venoms. Comparison of protein compositions of O.hannah venoms were performed using 1-D gel electrophoresis, 2D-gel electrophoresis and reverse-phase HPLC. Although protein compositions in all samples were similar, KV2 venom is very different from each other venom. The protein bands of KV2 in SDS-PAGE showed higher amount of all bands. 2DE gel of KV2 has many low molecular weight (14 kDa) spots. Spot No.2 and spot No.4 of KV2 are individual which are not found in KV6 and KV9. From peptides mass mapping by MALDI-TOF MS and amino acid sequence database searching, the amino acid sequence of protein spot 2 and spot 4 are similar to partial amino acid residue of phospholipase group from O.hannah venom. The RP-HPLC pattern of KV2 is different from other venoms. The eluted in range 31-50% ACN have high protein concentration than other venoms in same range. All of results indicated that variable protein composition were cause of difference in toxicity. The amino acid sequence of fraction 1.2.2 and 1.2.3, which were separated by ion exchange chromatography and gel filtration chromatography, were similar to partial amino acid residue of natrin and ophanin from Naja atra and O.hannah venom respectively.