Heterologous expressed toxic and non-toxic peptide variants of toxin CssII are capable to produce neutralizing antibodies against the venom of the scorpion Centruroides suffusus suffusus.
Two toxic and one non-toxic recombinant peptide variants of the mammalian neurotoxin CssII was cloned into the expression vector pQE30 containing a 6His-tag and a Factor Xa proteolytic cleavage site. The toxic recombinant peptides rCssII, HisrCssII and the non-toxic rCssIIE15R were expressed under induction with isopropyl thiogalactoside (IPTG), isolated using chromatographic techniques and folded correctly in vitro. The three recombinant variants showed similar secondary structures as the native CssII, but only the rCssIIE15R was not toxic to mice at concentrations up to 30microg/20g mouse body weight when injected intraperitoneally. All three recombinant peptides were capable of displacing the native CssII from their receptor sites in rat brain synaptosomes, suggesting that they had similar structural and functional characteristics of the native peptides. The three recombinant variants of CssII and the native one were used as antigens for immunization of New Zealand rabbits. The antibodies present in the rabbit antisera were able to recognize the native CssII. Additionally and more importantly, the sera of the immunized rabbits were able to neutralize both the native toxin CssII and the whole soluble venom of the scorpion Centruroides suffusus suffusus. These results indicate that the recombinant peptides can be used to produce antidotes against the venom of this species of scorpion.
Hernández-Salgado
Kenya
K
Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, UNAM. Apartado Postal 510-3, Cuernavaca, Morelos 61500, Mexico.
Estrada
Georgina
G
Olvera
Alejandro
A
Coronas
Fredy I
FI
Possani
Lourival D
LD
Corzo
Gerardo
G
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D001921:Brain / Q000378:metabolism* / Q000648:ultrastructure; D003001:Cloning, Molecular; D006801:Humans; D007114:Immunization; D007274:Injections, Intraperitoneal; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D012604:Scorpion Venoms / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D012605:Scorpions; D013574:Synaptosomes / Q000378:metabolism
D000906:Antibodies; D010446:Peptide Fragments; D012604:Scorpion Venoms; C066386:scorpion toxin Css II