Cloning and purification of alpha-neurotoxins from king cobra (Ophiophagus hannah).
Thirteen complete and three partial cDNA sequences were cloned from the constructed king cobra (Ophiophagus hannah) venom gland cDNA library. Phylogenetic analysis of nucleotide sequences of king cobra with those from other snake venoms revealed that obtained cDNAs are highly homologous to snake venom alpha-neurotoxins. Alignment of deduced mature peptide sequences of the obtained clones with those of other reported alpha-neurotoxins from the king cobra venom indicates that our obtained 16 clones belong to long-chain neurotoxins (seven), short-chain neurotoxins (seven), weak toxin (one) and variant (one), respectively. Up to now, two out of 16 newly cloned king cobra alpha-neurotoxins have identical amino acid sequences with CM-11 and Oh-6A/6B, which have been characterized from the same venom. Furthermore, five long-chain alpha-neurotoxins and two short-chain alpha-neurotoxins were purified from crude venom and their N-terminal amino acid sequences were determined. The cDNAs encoding the putative precursors of the purified native peptide were also determined based on the N-terminal amino acid sequencing. The purified alpha-neurotoxins showed different lethal activities on mice.
Department of Animal Toxinology, Kunming Institute of Zoology, The Chinese Academy of Sciences, 32 East Jiao Chang Road, Kunming 650223, Yunnan, China.
Ying-Ying
YY
He
Wei-Hui
WH
Lee
Yun
Y
Zhang
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001681:Biological Assay; D002681:China; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D016000:Cluster Analysis; D018076:DNA, Complementary / Q000235:genetics; D004546:Elapid Venoms / Q000235:genetics* / Q000633:toxicity; D017815:Elapidae; D007928:Lethal Dose 50; D051379:Mice; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000235:genetics* / Q000633:toxicity; D010802:Phylogeny; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D004546:Elapid Venoms; D009498:Neurotoxins