Received: November 22, 2008.

Accepted: September 15, 2009.

Abstract published online: October 9, 2009.

Full paper published online: November 30, 2009.

CLONING OF A NOVEL ACIDIC PHOSPHOLIPASE A2 FROM THE VENOM GLAND OF Crotalus durissus cascavella (BRAZILIAN NORTHEASTERN RATTLESNAKE)

Guarnieri MC (1), Melo ESL (1), Melo KMS (2), Albuquerque-Modesto JC (3), Prieto-da-Silva ARB (4), Rádis-Baptista G (2, 5)

(1) Department of Zoology, Federal University of Pernambuco, UFPE, Recife, Pernambuco State, Brazil; (2) Department of Biochemistry, Federal University of Pernambuco, UFPE, Recife, Pernambuco State, Brazil; (3) Department of Biology, Federal University of Pernambuco, UFPE, Vitória de Santo Antão, Pernambuco State, Brazil; (4) Laboratory of Genetics, Butantan Institute, São Paulo, São Paulo State, Brazil; (5) Institute of Marine Sciences, Federal University of Ceará, UFC, Fortaleza, Ceará State, Brazil.

ABSTRACT: The phospholipase A2 superfamily encompasses 15 groups that are classified into: secreted PLA2 (sPLA2); cytosolic PLA2 (cPLA2); Ca2+-independent intracellular PLA2 (iPLA2); platelet-activating factor acetylhydrolase (PAF-AH); and lysosomal PLA2. Currently, approximately 700 PLA2 sequences are known, of which 200 are obtained from the venom gland of Crotalinae snakes. However, thus far, little information is available on cloning, purification and structural characterization of PLA2 from Crotalus durisssus cascavela venom gland. In the present work, we report the molecular cloning of a novel svPLA2 from C. d. cascavella (Cdc), a predominant rattlesnake subspecies in northeastern Brazil. The Cdc svPLA2 cDNA precursor is 689 nucleotides long and encodes a protein of 138 amino acid residues, with a calculated molecular mass of approximately 13,847 Da and an estimated isoelectric point of 5.14. Phylogenetic analysis of Crotalinae PLA2 reveals that Cdc PLA2 clustered with other acidic type IIA PLA2 homologues is also present in the venom of North American rattlesnakes. Hitherto, this study presents a novel PLA2 cDNA precursor from C. d. cascavella and data reported herein will be useful for further steps in svPLA2 purification and analysis.

KEY WORDS: molecular toxinology, Crotalus durissus cascavella, snake venom gland, cDNA library, acidic PLA2.

CONFLICTS OF INTEREST: There is no conflict.

CORRESPONDENCE TO:

GHANDI RÁDIS-BAPTISTA, Instituto de Ciências do Mar, Labomar, Universidade Federal do Ceará, Av. da Abolição, 3207, Fortaleza, CE, 60165-081, Brasil. Phone: +55 85 3366 7000. Fax: +55 85 3242 8355. Email: gandhi.radis@ufc.br.