Effect of rACLF, a recombinant snake venom metallopeptidase on cell viability, chemokine expression and degradation of extracellular matrix proteins.
Snake venom metallopeptidases (SVMPs) comprise a family of zinc-dependent enzymes, which display many different biological activities. ACLF is a 23kDa fibrinolytic non-hemorrhagic metallopeptidase from the venom of the snake Agkistrodon contortrix laticinctus. We have previously developed an expression system for production of recombinant ACLF (rACLF) in bacteria. To achieve a better understanding of the role of such enzyme in envenoming cases, we have studied the biological properties of rACLF, including the ability of enzyme to degrade extracellular proteins, as well its cytotoxic effect in human fibroblasts and HeLa cells. Our results showed that rACLF hydrolyzed laminin, fibronectin, type IV collagen and thrombospondin. rACLF decreased HeLa cell viability, changed cell morphology and induced detachment, while for human fibroblasts no cytotoxic effects were observed after treatment with rACLF. In addition, growth-related oncogene (GRO) and monocyte chemoattractant protein 1 (MCP-1/CCL2) were chemokines detected in the culture supernatant of human fibroblasts incubated with rACLF for 48h. These chemokines could contribute to the severe local lesion induced by Agkistrodon contortrix lacticinctus venom. These findings suggest a relevant role for ACLF in envenomation.
de Moraes
Caroline Krieger
CK
Departamento de Ciências Fisiológicas, Universidade Federal de São Carlos, Rodovia Washington Luís Km 235, São Carlos, SP 13565-905, Brazil.
Selistre-de-Araujo
Heloisa Sobreiro
HS
D017836:Agkistrodon / Q000378:metabolism*; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D018925:Chemokines / Q000378:metabolism*; D003435:Crotalid Venoms / Q000201:enzymology*; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis / Q000187:drug effects; D045726:Metalloproteases / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology*
D018925:Chemokines; D003435:Crotalid Venoms; D016326:Extracellular Matrix Proteins; D011994:Recombinant Proteins; D045726:Metalloproteases