Cloning, expression and characterization of a phospholipase D from Loxosceles gaucho venom gland
| Autor: | Maria C Caporrino, José Antonio Portes-Junior, Maisa S. Della-Casa, Louise F. Kimura, Daniel A. Fukuda, José Pedro Prezotto-Neto, Katia C. Barbaro, Geraldo S. Magalhães, Marcelo L. Santoro, Ana G.C. Neves-Ferreira |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Platelet Aggregation
medicine.drug_class Molecular Sequence Data Antivenom Gene Expression Spider Venoms Venom Cross Reactions Biology Monoclonal antibody medicine.disease_cause Hemolysis complex mixtures Biochemistry Microbiology Structure-Activity Relationship Phospholipase D medicine Animals Humans Amino Acid Sequence Cloning Molecular Envenomation Base Sequence Toxin General Medicine medicine.disease Antibodies Neutralizing Loxoscelism Sphingomyelin Phosphodiesterase Immunology biology.protein Rabbits Antibody Sequence Alignment |
| Zdroj: | Biochimie. 95:1773-1783 |
| ISSN: | 0300-9084 |
| DOI: | 10.1016/j.biochi.2013.06.002 |
| Popis: | 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 (~65%) and dermonecrosis (~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. |
| Databáze: | OpenAIRE |
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