title: Isolation and expression of a hypotensive and anti-platelet acidic phospholipase A2 from Bothrops moojeni snake venom.
abstract: Phospholipases A(2) are important components of snake venoms, the basic isoforms have been more extensively studied than the acidic groups, maybe due to their higher toxicity. Trying to better understand the role of the acidic isoforms on the envenomation process, an acidic phospholipase A(2) was purified from Bothrops moojeni snake venom through two chromatographic steps (BmooPLA(2)). The enzyme showed a relative molecular mass of 13,601Da, pI 5.2, high phospholipase activity, bactericidal effect, moderate cytotoxic activity and was able to inhibit platelet aggregation. Moreover, BmooPLA(2) induced moderate in vivo edema and hypotensive effect. The 414bp cDNA encoding the BmooPLA(2) was cloned and expressed in Escherichia coli. The recombinant BmooPLA(2) showed phospholipase and inhibitory activities on platelet aggregation similar to those of the native protein. A comparative study between BmooPLA(2), the acidic (BthA-I) and basic (BthTX-II) PLA(2) from B. jararacussu venom showed that the effects of BmooPLA(2) and BthA-I-PLA(2) are similar. BmooPLA(2) is the first isolated and characterized non-myotoxic PLA(2) from B. moojeni snake venom. The recombinant PLA(2) can substitute the native toxin in studies aiming its biotechnological application in order to help the preservation of this endangered species. These data along with the preliminary structural studies here reported will provide a better understanding of this important class of proteins.
authors.affiliation: Departamento de Física e Química, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto-SP, Brazil.
authors.forename: Lucas B
authors.initials: LB
authors.lastname: Silveira
authors.forename: Daniela P
authors.initials: DP
authors.lastname: Marchi-Salvador
authors.forename: Norival A
authors.initials: NA
authors.lastname: Santos-Filho
authors.forename: Floriano P
authors.initials: FP
authors.lastname: Silva
authors.forename: Silvana
authors.initials: S
authors.lastname: Marcussi
authors.forename: André L
authors.initials: AL
authors.lastname: Fuly
authors.forename: Auro
authors.initials: A
authors.lastname: Nomizo
authors.forename: Saulo L
authors.initials: SL
authors.lastname: da Silva
authors.forename: Rodrigo G
authors.initials: RG
authors.lastname: Stábeli
authors.forename: Eliane C
authors.initials: EC
authors.lastname: Arantes
authors.forename: Andreimar M
authors.initials: AM
authors.lastname: Soares
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000302:isolation & purification / Q000494:pharmacology; D000959:Antihypertensive Agents / Q000302:isolation & purification* / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000302:isolation & purification / Q000494:pharmacology; D017837:Bothrops; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000737:chemistry* / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008297:Male; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D054467:Phospholipases A2 / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects; D010975:Platelet Aggregation Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D011487:Protein Conformation; D011817:Rabbits
chemical_list: D000900:Anti-Bacterial Agents; D000959:Antihypertensive Agents; D000970:Antineoplastic Agents; D003435:Crotalid Venoms; D010975:Platelet Aggregation Inhibitors; D054467:Phospholipases A2