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Ontogenetic variations in the venom proteome of the Amazonian snake Bothrops atrox.

Guércio RA, Shevchenko A, Shevchenko A, López-Lozano JL, Paba J, Sousa MV, Ricart CA - Proteome Sci (2006)

Bottom Line: The 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.Our findings underscore the importance of the use of venoms from individual specimen at various stages of maturation for the production of antivenoms.

View Article: PubMed Central - HTML - PubMed

Affiliation: Brazilian Center for Protein Research, Department of Cell Biology, University of Brasilia, Brasília, 70910-900- DF, Brazil. raffa@unb.br

ABSTRACT

Background: Bothrops 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.

Results: Optimized 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.

Conclusion: The 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.

No MeSH data available.


Related in: MedlinePlus

Proteome maps of B. atrox venom from juveniles, sub-adults and adults. Spots displaying similar peptide mass fingerprints were grouped as explained in Results section. A summary of the protein classes identified in each group is also shown.
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Figure 1: Proteome maps of B. atrox venom from juveniles, sub-adults and adults. Spots displaying similar peptide mass fingerprints were grouped as explained in Results section. A summary of the protein classes identified in each group is also shown.

Mentions: Patterns of protein spots visualized by silver staining were different between pooled venom samples from juvenile, sub-adult and adult B. atrox (Fig. 1). Their computer-assisted image analyses detected 110 spots in the gels from juveniles, 101 in sub-adults and 86 in adult venoms. Among the detected spots, 44 were found specifically in juveniles, 22 in sub-adults and 22 in adults. Image analysis also pinpointed substantial differences in the relative abundance of several spots matched at all three images (Table 1).


Ontogenetic variations in the venom proteome of the Amazonian snake Bothrops atrox.

Guércio RA, Shevchenko A, Shevchenko A, López-Lozano JL, Paba J, Sousa MV, Ricart CA - Proteome Sci (2006)

Proteome maps of B. atrox venom from juveniles, sub-adults and adults. Spots displaying similar peptide mass fingerprints were grouped as explained in Results section. A summary of the protein classes identified in each group is also shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1483819&req=5

Figure 1: Proteome maps of B. atrox venom from juveniles, sub-adults and adults. Spots displaying similar peptide mass fingerprints were grouped as explained in Results section. A summary of the protein classes identified in each group is also shown.
Mentions: Patterns of protein spots visualized by silver staining were different between pooled venom samples from juvenile, sub-adult and adult B. atrox (Fig. 1). Their computer-assisted image analyses detected 110 spots in the gels from juveniles, 101 in sub-adults and 86 in adult venoms. Among the detected spots, 44 were found specifically in juveniles, 22 in sub-adults and 22 in adults. Image analysis also pinpointed substantial differences in the relative abundance of several spots matched at all three images (Table 1).

Bottom Line: The 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.Our findings underscore the importance of the use of venoms from individual specimen at various stages of maturation for the production of antivenoms.

View Article: PubMed Central - HTML - PubMed

Affiliation: Brazilian Center for Protein Research, Department of Cell Biology, University of Brasilia, Brasília, 70910-900- DF, Brazil. raffa@unb.br

ABSTRACT

Background: Bothrops 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.

Results: Optimized 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.

Conclusion: The 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.

No MeSH data available.


Related in: MedlinePlus