title: Inhibition of crotoxin binding to synaptosomes by a receptor-like protein from Crotalus durissus terrificus (the South American rattlesnake).
abstract: Crotoxin (Ctx) is a potent neurotoxin of the venom of Crotalus durissus terrificus (the South American rattlesnake). Ctx is a heterodimer composed of CB, a toxic PLA(2) subunit, and CA, a non-toxic and non-enzymatic subunit, that potentiates the neurotoxicity of CB in vivo. The deleterious action of Ctx upon C. d. terrificus snakes themselves is known to be prevented by a PLA(2) inhibitor (CNF) present in their blood serum. CNF acts by replacing CA in Ctx, thus forming a new stable complex CNF-CB. This complex no longer interacts with the target receptor (TR) to deliver CB to cause its lethal effect. Furthermore, CNF-CB seems to be reminiscent of the interaction Ctx-TR at the pre-synaptic site. In the present work, the binding competition between rat brain synaptosomes (TR) and CNF for Ctx was investigated. Radiolabeled Ctx, made of CA and one isoform of CB (CA-(125)ICB(2)), was used as ligand. The competition by unlabeled Ctx was taken as a reference. The potency of CNF as a competitor was evaluated under different incubation conditions with varying time scale addition of reagents (CA-(125)ICB(2), synaptosomes and CA-CB(2) or CNF). CNF was able to inhibit the binding of the toxin to synaptosomes as well as to partially displace the toxin already bound to its membrane target. The mechanisms of competition involved were discussed and a previous schematic model of interactions between Ctx, TR and CNF was updated.
authors.affiliation: Centro de Pesquisa e Desenvolvimento, Fundação Ezequiel Dias (FUNED), Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Roberta Márcia Marques
authors.initials: RM
authors.lastname: dos Santos
authors.forename: Leida Calegário
authors.initials: LC
authors.lastname: Oliveira
authors.forename: Maria Inácia
authors.initials: MI
authors.lastname: Estevão-Costa
authors.forename: Maria Elena
authors.initials: ME
authors.lastname: de Lima
authors.forename: Marcelo Matos
authors.initials: MM
authors.lastname: Santoro
authors.forename: Consuelo Latorre
authors.initials: CL
authors.lastname: Fortes-Dias
mesh_terms: D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D001921:Brain / Q000166:cytology; D003435:Crotalid Venoms / Q000037:antagonists & inhibitors / Q000378:metabolism; D017839:Crotalus; D003439:Crotoxin / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D051381:Rats; D011956:Receptors, Cell Surface / Q000187:drug effects* / Q000378:metabolism; D030162:Reptilian Proteins / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D013574:Synaptosomes / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*
chemical_list: D001798:Blood Proteins; D003435:Crotalid Venoms; D006023:Glycoproteins; C400481:PLIalpha; D021122:Protein Subunits; D011956:Receptors, Cell Surface; D030162:Reptilian Proteins; C087438:phospholipase A2 inhibitor protein, Crotalus; D003439:Crotoxin