Identification,cloning and functional characterization of a novel dermonecrotic toxin (phospholipase D) from brown spider (Loxosceles intermedia) venom |
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Authors: | Marcia Helena Appel Rafael Bertoni da Silveira Olga Meiri Chaim Kátia Sabrina Paludo Dilza Trevisan Silva Daniele M Chaves Paulo Henrique da Silva Oldemir C Mangili Andrea Senff-Ribeiro Waldemiro Gremski Helena B Nader Silvio Sanches Veiga |
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Institution: | 1. Department of Cell Biology, Federal University of Paraná, Jardim das Américas, 81531-990, Curitiba, Paraná, Brazil;2. Department of Biochemistry, Federal University of São Paulo, São Paulo, Brazil;3. Department of Physiology, Federal University of Paraná, Brazil;4. Catholic University of Paraná, Health and Biological Sciences Institute, Curitiba, Paraná, Brazil |
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Abstract: | Brown spider bites are associated with lesions including dermonecrosis, gravitational spreading and a massive inflammatory response, along with systemic problems that may include hematological disturbances and renal failure. The mechanisms by which the venom exerts its noxious effects are currently under investigation. It is known that the venom contains a major toxin (dermonecrotic toxin, biochemically a phospholipase D) that can experimentally induce dermonecrosis, inflammatory response, animal mortality and platelet aggregation. Herein, we describe cloning, heterologous expression, purification and functionality of a novel isoform of the 33 kDa dermonecrotic toxin. Circular dichroism analysis evidenced correct folding for the toxin. The recombinant toxin was recognized by whole venom serum antibodies and by a specific antibody to a previously described dermonecrotic toxin. The identified toxin was found to display phospholipase activity and dermonecrotic properties. Additionally, the toxin caused a massive inflammatory response in rabbit skin dermis, evoked platelet aggregation, increased vascular permeability, caused edema and death in mice. These characteristics in combination with functional studies for other dermonecrotic toxins illustrate that a family of dermonecrotic toxins exists, and includes a novel member with high activity that may be useful for future structural and functional studies. |
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Keywords: | Dermonecrotic toxin Phospholipase D Sphingomyelinase Loxosceles Venom |
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