Venom constituents of Notechis scutatus scutatus (Australian tiger snake) from differing geographic regions

Column chromatography and polyacrylamide gel electrophoresis of Notechis scutatus scutatus venom showed that the venoms from different geographical locations had variations in their constituents. The venom collected from South Australia region contained both notexin and notechis II-5. The relative quantity of notechis II-5 was about three times that of notexin. On the other hand, the venom from Victoria region contained large amounts of notexin, but lacked notechis II-5. Instead, an unknown nontoxic protein, designated as notechis II-5b, exhibiting weak phospholipase A2 activity appeared in the position of notechis II-5 elution. This protein had an N-terminal sequence of N-L-I-Q-L-S-N-M-I-K-C-A-I-P-G-S-Q-P-L-F, sharing 45% homology with notexin and notechis II-5 and 60% homology with notechis II-1. The antibodies raised against Trp-modified notexin inhibited the enzymatic activities of notexin and notechis II-5 by 88 and 68%, respectively. However, the affinity of notexin for the antibodies was nine-fold greater than that of notechis II-5. This result is contrary to the previous finding (Mollier et al., FEBS Lett. 250, 479-482, 1989) in which notexin and notechis II-5 had similar binding affinities for antibodies raised against native notexin. This observation suggests that the antibodies prepared in this study could differentiate between isoforms of notexin.