Phylogenetic Relationship of Endogenous Alpha-PLA2 Inhibitors from the Plasma of Snakes

¹ESTEVÃO-COSTA, MI, ²MACIEL, R, ²VELARDE,DT, ³FRANCO, GR, ¹FORTES-DIAS,CL

1 Centro de Pesquisa e Desenvolvimento e 2 Divisão de Imunobiológicos, Fundação Ezequiel Dias (FUNED), Belo Horizonte, MG; 3 Depto. de Bioquimica e Imunologia, UFMG, Belo Horizonte.

Several authors have reported the presence of endogenous PLA2 inhibitors in the blood plasma of snakes. According to their structure and specificity, these inhibitors were divided into three classes: alpha, beta and gamma. In the present work we investigated the presence of cDNA encoding alpha PLA2 inhibitors, structurally characterized by the presence of motifs similar to the carbohydrate recognition domain (CRD) of C-type lectins. Total RNA was extracted from the liver of eight venomous snake species, commonly found in Brazil: Crotalus durissus terrificus, Lachesis muta, Bothrops alternatus, Bothrops erythromelas, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni and Bothrops neuwiedi. cDNA was obtained by RT-PCR and DNA amplification was performed by PCR in the presence of specific oligonucleotides designed on basis of the published nucleotide sequence of an alpha-inhibitor. The amplification products were cloned in a plasmid vector and the recombinant sequences obtained were aligned with the sequences of known alpha-inhibitors, using the Clustal W algorithm. A phylogenetic tree was derived for the members of this class of inhibitors.

FINANCIAL SUPPORT: FAPEMIG and CNPq

Correspondence to: mina@funed.mg.gov.br