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23 Congress of the International Union for Biochemistry and Molecular Biology
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44 Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology
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Foz do Iguaçu, PR, Brazil, August 24 to 28 , 2015
AN APPROACH TO THE SIALOME OF THE URUGUAYAN
SNAKE, BOTHROPS PUBESCENS (SERPENTES: VIPERIDAE)
Berasain, P.1; Carreira, S.2,3; Staats, C.C. 4; Termignoni, C. 5; Morais, V.6
1
Unidad Biología Parasitaria, Facultad de Ciencias, Instituto de Higiene,
Universidad de la República, Montevideo, Uruguay
2
Bioterio de Animales Ponzoñosos (Serpentario), Instituto de Higiene, UdelaR,
Montevideo, Uruguay
3
Laboratorio de Sistemática e Historia Natural de Vertebrados, Instituto de
Ecología y Ciencias Ambientales, Facultad de Ciencias, Universidad de la
República, Montevideo, Uruguay
4
Unidade de Química de Proteínas e Espectrometria de Massas, Centro de
Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil
5
Centro de Biotecnologia e Departamento de Bioquímica Universidade Federal
do Rio Grande do Sul, Porto Alegre, Brazil
6
Departamento de Desarrollo Biotecnológico, Fac. de Medicina, Instituto de
Higiene, Universidad de la República, Montevideo, Uruguay
INTRODUCTION AND OBJECTIVES: In Uruguay, 32-84 snakebites accidents
are reported each year (mean = 58, from 1514 cases in 27 years), caused by
two snakes included in the genus Bothrops: B. alternatus and B. pubescens.
The later belonged to the neuwiedi complex and is distributed in Uruguay and
southern Rio Grande do Sul, Brazil. Snake venom is a highly developed form of
saliva. In this work we propose to investigate the composition of the
venoms/saliva and find similarities and differences with other Bothrops
sialomes. MATERIALS AND METHODS: The experimental strategy consisted
of 1D and 2D gel electrophoresis analysis of a mixture of crude venom from
several B. pubescens snakes from serpentarium (Instituto de Higiene), excised
protein spots subjected to trypsin digestion and peptides analyzed by LCMS/MS mass spectrometry. Mass data were analyzed against public sequence
banks by using the Mascot search service. Complementary, biochemical assays
were performed in order to confirm the activity of the identified toxins.
RESULTS AND CONCLUSIONS: More than 70% of analyzed proteins in gel
were identified using Mascot public files. The main toxins reported to other
Bothrops venoms were also present in the venom of these Uruguayan snakes:
metalloproteinases, serinoproteinases, phospholipases and C-type lectin
proteins. The most abundant family seems to be the phospholipases which
performs more than 35% of the total amount of the venom. Phospholipase
activity measured by indirect hemolysis was compared between both species;
resulting B. pubescens venom has ten times higher activity than B. alternatus,
which represents the most important difference between the two Bothrops
species living in Uruguay. This is the first report for a proteome of B. pubescens
venom.
Acknowledgements: DICYT- CNPq, PEDECIBA QUÍMICA, CSIC
Keywords: Bothrops pubescens, Uruguay, Venoms.
Brazilian Society for Biochemistry and