Hindawi Publishing Corporation
BioMed Research International
Volume 2013, Article ID 591470, 13 pages
http://dx.doi.org/10.1155/2013/591470
Research Article
Biochemical Characterization and Pharmacological Properties of
New Basic PLA
BrTX-I Isolated from Bothrops roedingeri
(Roedinger’s Lancehead) Mertens, 1942, Snake Venom
Mauricio Aurelio Gomes Heleno, Paulo Aparecido Baldasso,
Luis Alberto Ponce-Soto, and Sérgio Marangoni
Department of Biochemistry, Institute of Biology, State University of Campinas (UNICAMP), P.O. Box 6109,
13083-970 Campinas, SP, Brazil
Correspondence should be addressed to Luis Alberto Ponce-Soto; poncesoto@yahoo.com.ar
Received 26 October 2012; Accepted 7 November 2012
Academic Editor: Elen Cristina Teizem Landucci
Copyright © 2013 Mauricio Aurelio Gomes Heleno et al. is is an open access article distributed under the Creative Commons
Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited.
BrTX-I, a PLA
2
, was puri�ed from Bothrops roedingeri venom aer only one chromatographic step using reverse-phase HPLC on
-Bondapak C-18 column. A molecular mass of 14358.69 Da was determined by MALDI-TOF mass spectrometry. Amino acid
analysis showed a high content of hydrophobic and basic amino acids as well as 14 half-cysteine residues. e total amino acid
sequence was obtained using SwissProt database and showed high amino acid sequence identity with other PLA
2
from snake venom.
e amino acid composition showed that BrTX-I has a high content of Lys, Tyr, Gly, Pro, and 14 half-Cys residues, typical of a basic
PLA
2
. BrTX-I presented PLA
2
activity and showed a minimum sigmoidal behavior, reaching its maximal activity at pH 8.0, 35–45
∘
C,
and required Ca
2+
. In vitro, the whole venom and BrTX-I caused a neuromuscular blockade in biventer cervicis preparations in
a similar way to other Bothrops species. BrTX-I induced myonecrosis and oedema-forming activity analyzed through injection of
the puri�ed BrTX-I in mice. Since BrTX-I exerts a strong proin�ammatory effect, the enzymatic phospholipid hydrolysis might
be relevant for these phenomena; incrementing levels of IL-1, IL-6, and TNF were observed at 15 min, 30 min, one, two, and six
hours postinjection, respectively.
1. Introduction
PLA
2
s (phosphatide 2-acylhydrolase, EC 3.1.14) represent a
superfamily of lipolytic enzymes which speci�cally catalyze
the hydrolysis of the ester bond at the sn-2 position of
glycerophospholipids resulting in the generation of fatty acid
(arachidonate) and lysophospholipids. e PLA
2
superfamily
consists of about 15 groups which are further subdivided into
several subgroups, all of which display differences in terms
of their structural and functional speci�cities. However,
the four main types or classes of PLA
2
s are the secreted,
the cytosolic, the Ca
2+
-independent and the lipoprotein-
associated PLA
2
[1], PLA
2
structure/function, mechanism,
and signaling [2].
Snake venom PLA
2
s displays a variety of activities, such
as neurotoxicity, myotoxicity, cardiotoxicity, and hemolysis
that may be modulated by speci�c receptors located on target
cells [3–6]. Indeed, PLA
2
receptors classi�ed as kinds M and
N [7] have been identi�ed in various kinds of cells, including
vascular smooth muscle cells, platelets, neutrophils, chondro-
cytes, �broblasts, hepatocytes, and mesangial cells, as well as
in brain, lung, and skeletal muscle [8, 9]. Snake venom PLA
2
can bind to M receptors, which are the most common kind
found in human macrophages and muscle cells, and these
may mediate some of the deleterious actions of venom PLA
2
s,
although that was not conclusively demonstrated [5, 6].
Peru has a rich and diverse herpetofauna that includes
venomous snake species of the families Elapidae (16 species