rd
23 Congress of the International Union for Biochemistry and Molecular Biology
th
44 Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology
th
th
Foz do Iguaçu, PR, Brazil, August 24 to 28 , 2015
MALE AND FEMALE SNAKE VENOM: DIFFERENCES IN PROTEIN
ISOFORMS AND CONCENTRATIONS.
Salvador, G.H.M.1, Sousa, I.D.L.2, Frihling, B.E.F.3, Santa-Rita, P.H.3, Fontes,
M.R.M.1, Marchi-Salvador, D.P.2
1
Departamento de Física e Biofísica, Instituto de Biociências, Universidade
Estadual Paulista, Botucatu, SP, Brazil; 2Departamento de Biologia
Molecular, Centro de Ciências Exatas e da Natureza, Universidade Federal
da Paraíba, João Pessoa, PB, Brazil; 3Biotério da Universidade Católica Dom
Bosco, Pró Reitoria de Pesquisa e Graduação, Universidade Católica Dom
Bosco, Campo Grande, MS, Brazil.
The Bothrops mattogrossensis is found in the eastern region of South America
including Bolivia, Brazil, Southeast Peru, Paraguay, Uruguay, and Argentina. The
envenomation by “boca de sapo”, as B. mattogrossensis is popularly known,
causes pain, oedema and bruising. This snake venom shows hemorrhagic,
platelet aggregation, proteolytic and myotoxic activities. The present study
describes the comparison between venom constituents from males and females of
B. mattogrossensis determined by Reverse-Phase Chromatography. Crude
venoms of B. mattogrossensis were pooled in two separated samples, specimens
from adult males and females from Mato Grosso do Sul kept in captivity at the
serpentarium of Dom Bosco Catholic University, Campo Grande, Brazil. For
Reverse-Phase HPLC separations, female and male lyophilized crude venom
were separate dissolved in 0.05% TFA and 5% acetonitrile, and insoluble material
were removed by centrifugation at 13,000×g for 10 min at 4°C. Proteins in the
soluble fraction were separated on a Shimadzu Shim-Pack C18 Column eluted at
1 mL/min with a linear gradient of 0.1% TFA in water (solution A) and 100% of
acetonitrile (solution B). The elution of absorbed proteins resulted in twenty-four
(female) or twenty (male) fractions. The chromatogram of the male and female
venom from Reverse-Phase Chromatography were similar, but the male venom
fraction eight (70-80 min) showed four weak peaks and the fraction ten (70-80
min) from female venom indicated only a concise peak. The molecular mass from
all HPLC fractions were performed by SDS-PAGE and the female fraction ten
measured around 13kDa. Although the female and male venom composition
showed similar, the female 13kDa fraction could not be observed in the male
venom. This fact indicated that males and females of the same species, kept in the
same environment with the same diet, produce venoms with different protein
isoforms and its concentrations.
Brazilian Society for Biochemistry and
Molecular Biology (SBBq)