Ontogenetic variations in the venom proteome of the Amazonian snake Bothrops atroxReportar como inadecuado




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Proteome Science

, 4:11

First Online: 11 May 2006Received: 14 December 2005Accepted: 11 May 2006

Abstract

BackgroundBothrops atrox is responsible for the majority of snakebite accidents in the Brazilian Amazon region. Previous studies have demonstrated that the biological and pharmacological activities of B. atrox venom alter with the age of the animal. Here, we present a comparative proteome analysis of B. atrox venom collected from specimens of three different stages of maturation: juveniles, sub-adults and adults.

ResultsOptimized conditions for two-dimensional gel electrophoresis 2-DE of pooled venom samples were achieved using immobilized pH gradient IPG gels of non-linear 3–10 pH range during the isoelectric focusing step and 10–20% gradient polyacrylamide gels in the second dimension. Software-assisted analysis of the 2-DE gels images demonstrated differences in the number and intensity of spots in juvenile, sub-adult and adult venoms. Although peptide mass fingerprinting PMF failed to identify even a minor fraction of spots, it allowed us to group spots that displayed similar peptide maps. The spots were subjected to a combination of tandem mass spectrometry and Mascot and MS BLAST database searches that identified several classes of proteins, including metalloproteinases, serine proteinases, lectins, phospholipases A2, L-amino oxidases, nerve growth factors, vascular endothelial growth factors and cysteine-rich secretory proteins.

ConclusionThe analysis of B. atrox samples from specimens of different ages by 2-DE and mass spectrometry suggested that venom proteome alters upon ontogenetic development. We identified stage specific and differentially expressed polypeptides that may be responsible for the activities of the venom in each developmental stage. The results provide insight into the molecular basis of the relation between symptomatology of snakebite accidents in humans and the venom composition. Our findings underscore the importance of the use of venoms from individual specimen at various stages of maturation for the production of antivenoms.

Abbreviations2-DEtwo-dimensional gel electrophoresis

IPGimmobilized pH gradient

CRISPCysteine Rich Secretory Proteins

PLA2phospholipase A2,

VEGFvascular endothelial growth factor

NGFnerve growth factor

IEFisoelectric focusing

DTTdithiotreitol

PMSFphenylmethyl sulfonyl fluoride

EDTAethylenediaminetetraacetic acid.

Electronic supplementary materialThe online version of this article doi:10.1186-1477-5956-4-11 contains supplementary material, which is available to authorized users.

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Fuente: https://link.springer.com/







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