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Title: Six isoforms of cardiotoxin in Malayan spitting cobra (Naja naja sputatrix) venom: Cloning and characterization of cDNAs
Authors: Jeyaseelan, K. 
Armugam, A. 
Lachumanan, R. 
Tan, C.H. 
Tan, N.H.
Keywords: (Naja naja sputatrix)
CDNA cloning
Issue Date: 10-Apr-1998
Citation: Jeyaseelan, K., Armugam, A., Lachumanan, R., Tan, C.H., Tan, N.H. (1998-04-10). Six isoforms of cardiotoxin in Malayan spitting cobra (Naja naja sputatrix) venom: Cloning and characterization of cDNAs. Biochimica et Biophysica Acta - General Subjects 1380 (2) : 209-222. ScholarBank@NUS Repository.
Abstract: Cardiotoxins are the most abundant toxin components of cobra venom. Although many cardiotoxins have been purified and characterized by amino acid sequencing and other pharmacological and biochemical studies, to date only five cardiotoxin cDNAs from Taiwan cobra (Naja naja atra), three cDNAs from Chinese cobra (Naja atra) and two more of uncertain origin (either Chinese or Taiwan cobra) have been reported. In this paper we show the existence of four isoforms of cardiotoxin by protein analysis and nine cDNA sequences encoding six isoforms of cardiotoxins (CTX 1-3, 4a, 4b and 5) from N.n. sputatrix by cDNA cloning. This forms the first report on the cloning and characterization of several cardiotoxin genes from a single species of a spitting cobra. The cDNAs encoding these isoforms, obtained by reverse transcription-polymerase chain reaction (RT-PCR), were subsequently expressed in Escherichia coli. The native and recombinant cardiotoxins were first characterized by Western blotting and N-terminal protein sequencing. These proteins were also found to have different levels of cytolytic activity on cultured baby hamster kidney cells. Four of the isoforms (CTX 1, 2, 4 and 5) are unique to N.n. sputatrix, with CTX 2 being the most abundant species constituting about 50% of the total cardiotoxins. The isoform CTX 3 (20% constitution) is highly homologous to the cardiotoxins of N.n. atra and N.n. naja, indicating that it may be universally present in all Naja naja subspecies. Our studies suggest that the most hydrophilic isoform (CTX 5) could have evolved first followed by the hydrophobic isoforms (CTX 1, 2, 3 and 4). We also speculate that Asiatic cobras could be the modem descendants of the African and Egyptian counterparts.
Source Title: Biochimica et Biophysica Acta - General Subjects
ISSN: 03044165
DOI: 10.1016/S0304-4165(97)00143-8
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