PHOSPHOLIPASIC ACTIVITY IN SKIN SECRETION OF AMAZONIAN ANURAN (AMPHIBIA) SPECIES

MARTINS M.J.P(1,3), ARAÚJO T.F(2,3), HIGA A. M(2,3), NORONHA M. D (2,3), MUNIZ E. G.(3), MEDEIROS B. M.(3, 4), LOPEZ-LOZANO J.L. (1,2,3)

(1) Programa de Pós-graduaçao em Biotecnologia - UFAM; (2) Mestrado em Biotecnologia - UEA; (3) Fundação de Medicina Tropical do Amazonas -FMTAM; (4) Curso em Ciências Biológicas UFAM.

Peptides (14KDa), alkaloids and steroids show several biological activities with biotechnological applications are principal compounds in anuran skin secretion. Skin secretion enzymes of Amazonian anuran species were not studied yet. The aim of this work was detect phospholipasic activity in skin secretion of amazonian anuran species. Specimens were collected in Manaus region and skin secretions were obtained by low voltage electric stimulation from dorsum skin glands. Skin secretions were lyophilized to be used in experiments. Protein molecular masses profiles of skin secretions was done using SDS-PAGE system. The phospholipasic activity was detected using 3% egg yolk incorporated in 1% agarose plate gel method in buffer PBS 0,10 mM pH 8,1 and 0,09 mM CaCl2. Immunological assays were performed with polyclonal antibodies against elapidae and crotalinae snake venoms. Bothrops atrox venom (10 µg) was used as phospholipase activity control. Phyllomedusa bicolor and P. tarsius showed phospholipasic activity. SDS-PAGE profile of both samples showed proteins bands between 24 kDa and 50 kDa range. Phospholipasic activity of P. tarsius sample (60 µg) was six times less potent that B. atrox venom and P. bicolor sample (90 µg) showed very low phospholipasic activity. Immunological tests showed no neutralization phospholipasic activity and non ELISA crosses reactions with polyclonal antibodies snake venons. The molecular characterization of proteins with phospholipasic activity is in course. Our data show new compounds in Amazonian anuran skin secretions with possible biotechnological relevance but without possible immunological relationships with elapidae and crotalinae toxins.

KEY WORDS: Amazonia, Anura, Skin Secretion, Phospholipase, Biotechnology.

FINANCIAL SUPPORT: CAPES, FMTAM, FINEP, FAPEAM

CORRESPONDENCE TO: MARX J. P. MARTINS, Centro de Ofidismo Prof. Paulo Friedrich Bürnheim/ FMTAM, Manaus, Brasil. Phone: + 55 92 3238 42 94. E-mail: batmartins@hotmail.com