title: A recombinant snake neurotoxin generated by chemical cleavage of a hybrid protein recovers full biological properties.
abstract: We previously reported the production of a fused snake neurotoxin composed of protein A and erabutoxin a in E. coli. The hybrid had much lower toxicity and affinity for the acetylcholine nicotinic receptor than natural erabutoxin. By treating the hybrid with cyanogen bromide we generated a toxin which was purified in a single step by RP-HPLC. This compound, produced in a good yield, recovered all properties of native erabutoxin a, implying that the lower toxic activities of the hybrid were due to the bulky protein A and not to an incorrect folding of the toxin. This work serves as a basis for future studies of toxin-receptor interactions using engineered toxin mutants.
authors.affiliation: Département de Biologie, Laboratoire d'Ingénierie des Protéines, C.E.N., Gif-sur-Yvette, France.
authors.forename: P
authors.initials: P
authors.lastname: Boyot
authors.forename: L
authors.initials: L
authors.lastname: Pillet
authors.forename: F
authors.initials: F
authors.lastname: Ducancel
authors.forename: J C
authors.initials: JC
authors.lastname: Boulain
authors.forename: O
authors.initials: O
authors.lastname: Tremeau
authors.forename: A
authors.initials: A
authors.lastname: Ménez
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000911:Antibodies, Monoclonal / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D003488:Cyanogen Bromide; D004546:Elapid Venoms / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000235:genetics* / Q000276:immunology; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A
chemical_list: D000596:Amino Acids; D000911:Antibodies, Monoclonal; D004546:Elapid Venoms; D009498:Neurotoxins; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D003488:Cyanogen Bromide