INHIBITORY PROPERTIES OF ANTI-BOTHROPIC COMPLEX FROM Didelphis albiventris SERUM AND POLYCLONAL ANTIBODIES ON PHARMACOLOGICAL EFFECTS OF TOXINS FROM Bothrops SNAKE VENOMS

SOARES, A.M.(1), TRENTO, E.P.(2), GIGLIO, J.R.(3)

(1)Departamento de Biotecnologia, UNAERP, Ribeirão Preto-SP, (2)Departamento de Bioquímica, UNIC, Cuiabá, MT and (3)Departamento de Bioquímica e Imunologia, FMRP, USP, Ribeirão Preto, SP.

Anti-bothropic complex (ABC) was isolated from the serum of the South American opossum (Didelphis albiventris) by single-step affinity chromatography using a Sepharose-immobilized metalloprotease (BaP1) from Bothrops asper as the binding protein. Functional studies pointed out that ABC inhibits the hemorrhagic and proteolyticactivities on fibrin, fibrinogen, and casein induced by the metalloproteasesisolated from B. asper BaP1 and BaH4, B. neuwiedi neuwiedase and B. moojeni MOO3. In addition to the anti-hemorrhagic and anti-proteolytic activities, ABC also showed anti-myotoxic, anti-lethal, and anti-edematogenic effects against myotoxicphospholipases A₂ isolated from B. asper Basp-I, II and III, B. moojeni MjTX-I and II, B. jararacussu BthTX-I and II, B. pirajai PrTX-I and III, and B. neuwiedi BnSP-7. Moreover, it had inhibitory effects on the phospholipase A₂ activity of the crude venom as well as the isolated venom phospholipasesA₂. Biochemical characterization of ABC showed the presence of two glycosylated subunits of 43 and 45 kDa, respectively, with an isoelectric point < 4. The two subunits were separated by ion-exchange HPLC. The N-terminal sequences of both subunits (LKAMDPTPXLWIETESP, where X is Arg-9 and Pro-9, respectively) showed a high degree of identity with other serum inhibitors isolated from different marsupials. Antibodies to ABC cross-reacted with different snake venoms, 04 isolated metalloproteasesand 12 purified class II myotoxicphospholipasesA₂ found in snake venoms of the genus Bothrops. In neutralization experiments, pre-incubation of crude venom or isolated toxins with antibodies to the ABC inhibited their hemorrhagic, PLA₂ and myotoxic activities.

Financial supported: FAPESP and CNPq.

CORRESPONDENCE TO:

SOARES, A.M., Departamento de Biotecnologia, UNAERP, Ribeirão Preto, SP, 14000-000, Brasil, Email: andreimar@unaerp.br