Cloning and characterization of an alpha-neurotoxin-type protein specific for the coral snake Micrurus corallinus.
During the cloning of abundant cDNAs expressed in the Micrurus corallinus coral snake venom gland, we cloned an alpha-neurotoxin homologue cDNA (nxh1). Two others isoforms were also cloned (nxh3 and nxh7, respectively). The nxh1 cDNA codes for a potential coral snake toxin with a signal peptide of 21 amino acids plus a predicted mature peptide with 57 amino acids. The deduced protein is highly similar to known toxic three-finger alpha-neurotoxins, with four deduced S-S bridges at the same conserved positions. This is the first cDNA coding for a three-finger related protein described so far for coral snakes. However, the predicted protein does not possess some of the important amino acids for the nicotinic acetylcholine receptor interaction. This protein was expressed in Escherichia coli as a His-tagged protein that allowed the rapid purification of the recombinant protein. This protein was used to generate antibodies which recognized the recombinant protein in Western blot and also a single band present in the M. corallinus venom, but not in the venom of 10 other Micrurus species.
Silveira de Oliveira
J
J
Centro de Biotecnologia, Instituto Butantan, São Paulo, SP, 05503-900, Brazil.
Rossan de Brandão Prieto da Silva
A
A
Soares
M B
MB
Stephano
M A
MA
de Oliveira Dias
W
W
Raw
I
I
Ho
P L
PL
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary; D004546:Elapid Venoms / Q000737:chemistry* / Q000235:genetics; D017815:Elapidae; D015723:Gene Library; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000737:chemistry*; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D012627:Sebaceous Glands / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D004546:Elapid Venoms; D009498:Neurotoxins; D020033:Protein Isoforms; D011994:Recombinant Proteins