Cloning, expression and characterization of a phospholipase D from Loxosceles gaucho venom gland

Autor(es)

Magalhaes, Geraldo S. ^[1] ; Caporrino, Maria C. ^[1] ; Della-Casa, Maisa S. ^[1] ; Kimura, Louise F. ^[1] ; Prezotto-Neto, Jose P. ^[1] ; Fukuda, Daniel A. ^[1] ; Portes-Junior, Jose A. ^[1] ; Neves-Ferreira, Ana G. C. ^[2] ; Santoro, Marcelo L. ^[3] ; Barbaro, Katia C. ^[1]

Afiliação do(s) autor(es)

^[1] Butantan Inst, Immunopathol Lab, BR-05503900 Sao Paulo - Brazil

^[2] Fiocruz MS, Inst Oswaldo Cruz, Lab Toxinol, BR-21040900 Rio De Janeiro, RJ - Brazil

^[3] Butantan Inst, Lab Pathophysiol, BR-05503900 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Biochimie; v. 95, n. 9, p. 1773-1783, SEP 2013.
• Citações Web of Science: 21

Resumo

Loxosceles venom comprises a mixture of diverse toxins that induces intense local inflammatory reaction, dermonecrotic injury, platelet aggregation, hemolytic anemia and acute renal failure. Among several toxins in the venom, phospholipases D (PLDs), also called dermonecrotic toxins, are the most important and best studied, since they account for the main effects observed in loxoscelism. Despite their importance, biological analysis of PLDs is hampered by the minute amounts normally purified from the venom, and therefore many efforts have been made to clone those toxins. However, to date, no PLD from Loxosceles gaucho has been obtained in a heterologous system. Thus, in this work we show the cloning of a PLD from L gaucho venom gland, named LgRec1, which was successfully expressed in a bacterial system. LgRec1 evoked local reaction (edema, erythema, ecchymosis, and paleness), dermonecrosis and hemolysis. It was also able to hydrolyze sphingomyelin and promote platelet aggregation. ELISA and Western blot analysis showed that LgRec1 was recognized by an anti-L gaucho venom serum, a commercial arachnidic antivenom as well as a monoclonal antibody raised against the dermonecrotic fraction of L gaucho venom. In addition, LgRec1 demonstrated to be highly immunogenic and antibodies raised against this recombinant toxin inhibited local reaction (similar to 65%) and dermonecrosis (similar to 100%) elicited by L gaucho whole venom. Since PLDs are considered the major components accounting for the local and systemic envenomation effects caused by spiders from genus Loxosceles, the information provided here may help to understand the mechanisms behind clinical symptomatology. (C) 2013 Elsevier Masson SAS. All rights reserved. (AU)

• Processo FAPESP: 11/23273-7 - Isolamento, clonagem e expressão de toxinas da glândula de veneno da aranha Loxosceles gaucho e interação com anticorpos
• Beneficiário: Katia Cristina Barbaro Nogueira
• Modalidade de apoio: Auxílio à Pesquisa - Regular