Two novel dermonecrotic toxins LiRecDT4 and LiRecDT5 from Brown spider (Loxosceles intermedia) venom: From cloning to functional characterization

Two novel dermonecrotic toxins LiRecDT4 and LiRecDT5 from Brown spider (Loxosceles intermedia) venom: From cloning to functional characterization

Author Silveira, Rafael Bertoni da Google Scholar
Pigozzo, Romine Bachmann Google Scholar
Chaim, Olga Meiri Google Scholar
Appel, Marcia Helena Google Scholar
Silva, Dilza Trevisan Google Scholar
Dreyfuss, Juliana Luporini Google Scholar
Toma, Leny Google Scholar
Dietrich, Carl Peter Google Scholar
Nader, Helena B. Google Scholar
Veiga, Silvio Sanches Google Scholar
Gremski, Waldemiro Google Scholar
Institution Univ Fed Parana
Universidade Federal de São Paulo (UNIFESP)
Catholic Univ Parana
Abstract Loxoscelism (the condition produced by the bite of brown spiders) has been reported worldwide, but especially in warmer regions. Clinical manifestations include skin necrosis with gravitational spreading while systemic loxoscelism may include renal failure, hemolysis and thrombocytopenia. the venom contains several toxins, of which the best biochemically and biologically studied is the dermonecrotic toxin, a phospholipase-D. Purified toxin induces cutaneous and systemic loxoscelism, especially necrotic lesions, hematological disturbances and renal failure. Herein, we describe cloning, heterologous expression and purification of two novel dermonecrotic toxins: LiRecDT4 and LiRecDT5. the recombinant proteins stably expressed in Escherichia coli cells were purified from culture supernatants in a single step using Ni(-)(2+)chelating chromatography producing soluble proteins of 34 kDa (LiRecDT4) and 37 kDa (LiRecDT5). Circular dichroism analysis evidenced correctly folding for toxins but differences in secondary structures. Both proteins were recognized by whole venom serum antibodies and by a specific antibody to dermonecrotic toxin. Also, recombinant toxins with phospholipase activity induced experimental skin lesions and caused a massive inflammatory response in rabbit skin dermis. Nevertheless, toxins displayed different effects upon platelet aggregation, increase in vascular permeability and not caused death in mice. These characteristics in combination with functional studies illustrates that a family of dermonecrotic toxins exists, and includes two novel members that are useful for future structural and functional studies. They will also be useful in biotechnological ends, for example, as inflammatory and platelet aggregating studies, as antigens for serum therapy source and for lipids biochemical research. (c) 2007 Elsevier Masson SAS. All rights reserved.
Keywords Brown spider
Loxosceles intermedia
venom
dermonecrotic toxin
phospholipase activity
Language English
Date 2007-03-01
Published in Biochimie. Paris: Elsevier France-editions Scientifiques Medicales Elsevier, v. 89, n. 3, p. 289-300, 2007.
ISSN 0300-9084 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 289-300
Origin http://dx.doi.org/10.1016/j.biochi.2006.12.002
Access rights Closed access
Type Article
Web of Science ID WOS:000245998900003
URI http://repositorio.unifesp.br/handle/11600/29541

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