Poster 116 - Partial proteomes of the venoms of spiders of the genera Phoneutria and Ancylometes

Poster 116. 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.475.

PARTIAL PROTEOMES OF THE VENOMS OF SPIDERS OF THE GENERA Phoneutria AND Ancylometes.

RICHARDSON, M.¹; PIMENTA, A.M.C.²; BEMQUERER, M.P.²; SANTORO, M.²; BEIRAO, P.S.L.²; LIMA, M.E.²; FIGUEIREDO, S.G.³; GOMES, P.C.¹; CORDEIRO, M.N.¹

1 Fundacao Ezequiel Dias, BH-MG, 2 Dept. Biochem. Immunol., UFMG, BH-MG, 3 Dept. Physiol. Sci.,UFES,Vitoria, ES.

The venoms of the Brazilian wandering “armed” spiders Phoneutrianigriventer, Ph reidyi, and Ph keyserling, and the South American fishing spider Ancylometes spp. were fractionated using a combination of preparative reverse phase HPLC on Vydac C4, analytical RV-HPLC on Vydac C8 and C18 and ion exchange FPLC on Resource S at pH 6.1 and 4.7. The homogeneity and molecular masses of the purified components were determined by high resolution propionic acid/urea PAGE and by electrospray MS (Q-TOF Micro) in positive ion mode. The amino acid sequences of the native and S-pyridylethylated proteins and peptides derived from them by digests with trypsin, chymotrypsin and/or a GLU-C protease were determined by Edman degradation using a Shimadzu PPSQ-21^A automated protein sequencer. The structures of some small peptides with modified N- and C-termini were obtained by analses of MS/MS collision induced dissociations.

To date nearly 400 peptides and proteins (1.2- 23kDa) have been isolated in a pure state, and of these more than 100 have had their complete or partial amino acid sequences determined. These sequences demonstrate, as might be expected, that the venoms of Ph. reidyi and Ph. keyserling (Family: Ctenidae) both contain a similar range of isoforms of the neurotoxins previously isolated from Ph. nigriventer which are active on neuronal ion (Ca²⁺, Na⁺ and K⁺) channels and NMDA- type glutamate receptors. However, somewhat surprisingly, the venom of Ancylometes (Family: Pisauridae) was also found to contain homologues of most of these types of molecules. The structures of a group of small (1.1-1.7kDa) peptides which cause contraction of smooth muscle were elucidated. In addition several new groups of structurally related polypeptides of as yet unknown function have also been identified. Two enzymes which hydrolyzed succinyl casein and gelatin were found in the venom of Ph. nigriventer and shown to have strong sequence identity with a peptide isomerase from the funnel web spider Agelenopsis and other type1 serine proteases. These enzymes may be responsible for the variety of examples of post-translational modifications observed in the sequences of some of the venom components.