Cloning and characterization of a basic phospholipase A2 homologue from Micrurus corallinus (coral snake) venom gland.
During the cloning of abundant cDNAs expressed in the Micrurus corallinus coral snake venom gland, several putative toxins, including a phospholipase A2 homologue cDNA (clone V2), were identified. The V2 cDNA clone codes for a potential coral snake toxin with a signal peptide of 27 amino acid residues plus a predicted mature protein with 119 amino acid residues. The deduced protein is highly similar to known phospholipases A2, with seven deduced S-S bridges at the same conserved positions. 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. This antiserum was used to screen a large number of venoms, showing a ubiquitous distribution of immunorelated proteins in all elapidic venoms but not in the viperidic Bothrops jararaca venom. This is the first description of a complete primary structure of a phospholipase A2 homologue deduced by cDNA cloning from a coral snake.
de Oliveira
Ursula Castro
UC
Centro de Biotecnologia, Instituto Butantan, Av. Vital Brazil, 1500, CEP:05503-900, São Paulo, SP, Brazil.
Assui
Alessandra
A
da Silva
Alvaro Rossan de Brandão Prieto
AR
de Oliveira
Jane Silveira
JS
Ho
Paulo Lee
PL
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics; D004546:Elapid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics*; D017815:Elapidae / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli; D008969:Molecular Sequence Data; D010741:Phospholipases A / Q000235:genetics* / Q000302:isolation & purification; D054467:Phospholipases A2; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D016415:Sequence Alignment; D017385:Sequence Homology
D018076:DNA, Complementary; D004546:Elapid Venoms; D011994:Recombinant Proteins; C053972:micrurus venom; D010741:Phospholipases A; D054467:Phospholipases A2