Poster 102 - Comparative studies of the composition of the venoms of male and female spiders of

Poster 102. Congresso da Sociedade Brasileira de Toxinologia, 8., Symposium of the Pan American Section of the International Society on Toxinology, 8., 2004, Angra dos Reis, Brasil. Abstracts... J. Venom. Anim. Toxins incl.Trop. Dis., 2004, 10, 3, p.461.

Comparative Studies of the Composition of the Venoms of Male and Female Spiders of the Species Phoneutria keyserling and Phoneutria nigriventer.

Viana, N.P¹.; Pimenta, A.M.C².; Gomes, P.C¹.;Figueiredo, S.G³.;Richardson, M¹.;Cordeiro, M.N¹.

¹Fundação Ezequiel Dias, BH-MG,² Dept. Biochem. Immunol., UFMG, BH-MG,³Dept. Physiol. Sci., UFES, Vitória- E.S.

The compositions of the venoms of male and female spiders of the species P. keyserling and P. nigriventer were compared with the objective of determining the primary structures and biological activities of sex-specific polypeptide components.The protein content of the crude venoms was determined by the procedure of Lowry et al (1951). The venoms were then subjected to SDS-PAGE (Laemmly),SDS-Tricina and propionic acid/urea to demonstrate any differences in their protein compositions. Afterwards these venoms were fractionated by RPC4-HPLC. The homogeneity/purity of the purified components was determined by high resolution propionic acid/urea and confirmed by ES-Q-TOF-MS in the positive ion mode, which also revealed the molecular masses of the components. The amino acid sequences of the sex-specific proteins were determined by automated protein sequencer (Shimadzu PPSQ 21). Bands of proteins which apperared to be sex specific were observed in the gel electrophoretic profiles. It was also possible to observe differences in the numbers, molecular weights and concentrations of specific proteins in the reverse phase HPLC profiles and mass spectra. The propionic acid/urea gels of P. keyserling, showed a particular band which was only present in the venom from male spiders. Similary the peak (27) observed in the profile of the RP- HPLC separation of the venoms of spiders of P. keyserling was much larger in the case of the venom from males. The predominant protein in this peak (male) was found to have identical amino acid sequence to the insecticidal toxin TX4 (5-5), previously isolated by us from the mixed venom (male and female) of spiders of the species P. nigriventer (Figueiredo et al, 2001). The venom from female spiders of P. keyserling also produced a peak (29) which seemed to be exclusive/specific. The results indicate that there are differences in the compositions of the venoms of male and female spiders of P. keyserling and P. nigriventer. Such differences in the compositions of sex specific venoms have been observed previously (Calegario et al 2001;Herzig et al 2002) but these workes only studied P. nigriventer and did not characterize or determine the structures of the sex specific proteins.