PROSPECTION OF VENOM PEPTIDASE ACTIVITIES VIA FLUORESCENCE-BASED ASSAY OF NAPHTHYLAMIDE DERIVATIVES

GASPARELLO-CLEMENTE, E.,  OLIVO, R.A.; SILVEIRA, P.F.

Laboratory of Pharmacology, InstitutoButantan

The fluorometry of naphthylamide derivatives was employed for revealing representative peptidase activities in whole venoms of the snakes Bothrops jararaca, Bothrops alternatus, Bothrops atrox, Bothrops moojeni, Bothrops insularis, Crotalus durissus terrificus and Bitis arietans, of the scorpion Tityus bahiensis, and of the spiders Phoneutria nigriventer and Loxosceles intermedia. As a result, neutral aminopeptidase (APN) and prolyl-dipeptidyl aminopeptidase IV (DPP IV) were presented in all snake venoms (highest levels in Bothrops alternatus). All examined peptidases showed relatively low levels in arthropod venoms, except basic aminopeptidase (APB) from Phoneutria nigriventer venom. Relatively high levels of acid aminopeptidase (APA) were restricted to Bitis arietans venom. Bitis arietans also exhibited a prominent content of APB which was lower in the other venoms. Prolyl endopeptidase and proline iminopeptidase activities were respectively detectable only at low levels in Tityus bahiensis and Bothrops insularis while pyroglutamate aminopeptidase activity was undetectable in all venoms. In conclusion, the difference between snake and arthropod venoms was mainly represented by APN and also by DPP IV activities. The contents of APA and APB activities constituted the difference between Bitis arietans and Brazilian snake venoms. The distribution of the peptidase activities investigated reflected an interesting trend of evolutionary divergence in the different processing of peptides in different venoms and/or in different abilities of the venoms examined to hydrolyze different peptides during envenomation. Our data also provide evidence that the measurement of venom peptidase activities may be useful for the purposes of ecophysiological studies, for taxonomic classification of venomous animals, and to elucidate the evolutive history of their venoms.

FAPESP-FUNDAP fellowship. Research Grant 00/10023-8 from FAPESP.

CORRESPONDENCE TO:

Renata do Amaral Olivo, Rua General Geronimo dos Reis , 452 A, São Paulo, 02211040, SP, Brasil, Email: renataolivo@hotmail.com