Venom gland EST analysis of the saw-scaled viper, Echis ocellatus, reveals novel α9β1 integrin-binding motifs in venom metalloproteinases and a new group of putative toxins, renin-like aspartic proteases

Wagstaff, Simon C.; Harrison, Robert A.

doi:10.1016/j.gene.2006.03.008

Cited by 97 publications

(76 citation statements)

References 52 publications

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“…These species were selected based on their medical importance in Africa and our previous descriptions of interspecific variation in venom composition, dietary preference, and prey lethality (12,20,25,26). We used our previously constructed and assembled venom gland transcriptomes for the four Echis species (26,27) and prepared venom gland trancriptomes for B. arietans and C. cerastes by using identical protocols. We generated proteomes from venom extracted from each of these species (28) and used translations of the transcriptomic datasets to facilitate protein identification (SI Appendix, Figs.…”

Section: Resultsmentioning

confidence: 99%

“…Venom gland transcriptomes for B. arietans (Nigeria) and C. cerastes (Egypt) were constructed using the protocols previously described for E. ocellatus (Nigeria), E. p. leakeyi (Kenya), E. coloratus (Egypt), and E. c. sochureki (United Arab Emirates) (26,27). Briefly, libraries were constructed by using mRNA extracted from venom glands pooled from 10 individuals of each species by using the CloneMiner method, with ∼1,000 clones sequenced by using Sanger sequencing for each library.…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, libraries were constructed by using mRNA extracted from venom glands pooled from 10 individuals of each species by using the CloneMiner method, with ∼1,000 clones sequenced by using Sanger sequencing for each library. Resulting ESTs were assembled into contigs (putative gene products) and annotated, and their transcription level was quantified as previously described (26,27). Each dataset was then subjected to six frame translations and used as a reference database to facilitate proteomic identification.…”

Section: Methodsmentioning

confidence: 99%

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Medically important differences in snake venom composition are dictated by distinct postgenomic mechanisms

Casewell

Wagstaff

Wüster

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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284

256

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Variation in venom composition is a ubiquitous phenomenon in snakes and occurs both interspecifically and intraspecifically. Venom variation can have severe outcomes for snakebite victims by rendering the specific antibodies found in antivenoms ineffective against heterologous toxins found in different venoms. The rapid evolutionary expansion of different toxin-encoding gene families in different snake lineages is widely perceived as the main cause of venom variation. However, this view is simplistic and disregards the understudied influence that processes acting on gene transcription and translation may have on the production of the venom proteome. Here, we assess the venom composition of six related viperid snakes and compare interspecific changes in the number of toxin genes, their transcription in the venom gland, and their translation into proteins secreted in venom. Our results reveal that multiple levels of regulation are responsible for generating variation in venom composition between related snake species. We demonstrate that differential levels of toxin transcription, translation, and their posttranslational modification have a substantial impact upon the resulting venom protein mixture. Notably, these processes act to varying extents on different toxin paralogs found in different snakes and are therefore likely to be as important as ancestral gene duplication events for generating compositionally distinct venom proteomes. Our results suggest that these processes may also contribute to altering the toxicity of snake venoms, and we demonstrate how this variability can undermine the treatment of a neglected tropical disease, snakebite.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Medically important differences in snake venom composition are dictated by distinct postgenomic mechanisms

Casewell

Wagstaff

Wüster

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

284

256

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“…less than 50 ng in 100 μg of venom proteins) to known protein families (Tables 1 and 2). Supporting the view that venom proteomes are mainly composed of toxins belonging to a few protein families [4,[9][10][11][12][13][14][16][17][18][19][20][21][22][23][24], the proteins found in the venoms of L. muta and L. stenophrys cluster, respectively, in 8 and 7 different families (bradykinin-potentiating peptides, NGF, PLA 2 , serine proteinase, cysteine-rich secretory proteins (CRISP; only found in L. muta), C-type lectins, L-amino acid oxidase (LAO), and Zn 2+ -dependent metalloproteinases) (Fig. 6), whose relative abundances are listed in Table 3.…”

Section: Characterization Of Bushmaster Venom Proteomesmentioning

confidence: 99%

“…Within-and between-species heterogeneity of venoms may also account for differences in the clinical symptoms observed in accidental envenomations. In order to explore the putative venom components, several laboratories have carried out transcriptomic analyses of the venom glands of viperid (Bitis gabonica [16], Bothrops insularis [17], Bothrops jararacussu [18], Bothrops jararaca [19], Agkistrodon acutus [20,21], Echis ocellatus [22], and Lachesis muta [23]), elapid (Oxyuramus scutellatus [24]), and colubrid (Philodryas olfersii [25]) snake species. Transcriptomic investigations provide catalogues of partial and full-length transcripts that are synthesized by the venom gland.…”

Section: Introductionmentioning

confidence: 99%

Snake venomics of the South and Central American Bushmasters. Comparison of the toxin composition of Lachesis muta gathered from proteomic versus transcriptomic analysis

Sanz

Escolano

Ferretti

et al. 2008

Journal of Proteomics

127

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This article appeared in a journal published by Elsevier. The attached copy is furnished to the author for internal non-commercial research and education use, including for instruction at the authors institution and sharing with colleagues.Other uses, including reproduction and distribution, or selling or licensing copies, or posting to personal, institutional or third party websites are prohibited. were identified in L. muta and L. stenophrys venoms. In addition, both venoms contained a large number of bradykinin-potentiating peptides (BPP) and a C-type natriuretic peptide (C-NP). BPPs and C-NP comprised around 15% of the total venom proteins. In both species, the most abundant proteins were Zn 2+ -metalloproteinases (32-38%) and serine proteinases (25-31%), followed by PLA 2 s (9-12%), galactose-specific C-type lectin (4-8%), L-amino acid oxidase (LAO, 3-5%), CRISP (1.8%; found in L. muta but not in L. stenophrys), and NGF (0.6%).On the other hand, only six L. muta venom-secreted proteins matched any of the previously reported 11 partial or full-length venom gland transcripts, and venom proteome and transcriptome depart in their relative abundances of different toxin families. As expected from their close phylogenetic relationship, the venoms of L. muta and L. stenophrys share (or contain highly similar) proteins, in particular BPPs, serine proteinases, a galactose-specific C-type lectin, and LAO. However, they dramatically depart in their respective PLA 2 complement. Intraspecific quantitative and qualitative differences in the expression of PLA 2 molecules were found when the venoms of five L. muta specimens (3 from Bolivia and 2 from Peru) and the venom of the same species purchased from Sigma were compared.These observations indicate that these class of toxins represents a rapidly-evolving Author's personal copy gene family, and suggests that functional differences due to structural changes in PLA 2 s molecules among these snakes may have been a hallmark during speciation and adaptation of diverging snake populations to new ecological niches, or competition for resources in existing ones. Our data may contribute to a deeper understanding of the biology and ecology of these snakes, and may also serve as a starting point for studying structure-function correlations of individual toxins.

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Snake venomics. Strategy and applications

2007

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Snake bites can be deadly, but the venoms also contain components of medical and biotechnological value. The proteomic characterization of snake venom proteomes, snake venomics, has thus a number of potential benefits for basic research, clinical diagnosis, and development of new research tools and drugs of potential clinical use. Snake venomics is also relevant for a deep understanding of the evolution and the biological effects of the venoms, and to generate immunization protocols to elicit toxin-specific antibodies with greater specificity and effectiveness than conventional systems. Our snake venomics approach starts with the fractionation of the crude venom by reverse-phase HPLC, followed by the initial characterization of each protein fraction by combination of N-terminal sequencing, SDS-PAGE, and mass spectrometric determination of the molecular masses and the cysteine (SH and S--S) content. Protein fractions showing a single electrophoretic band, molecular mass, and N-terminal sequence can be straightforwardly assigned by BLAST analysis to a known protein family. On the other hand, protein fractions showing heterogeneous or blocked N-termini are analyzed by SDS-PAGE and the bands of interest subjected to automated reduction, carbamidomethylation, and in-gel tryptic digestion. The resulting tryptic peptides are then analyzed by MALDI-TOF mass fingerprinting followed by amino acid sequence determination of selected doubly and triply charged peptide ions by collision-induced dissociation tandem mass spectrometry. The combined strategy allows us to assign unambiguously all the isolated venom toxins representing over 0.05% of the total venom proteins to known protein families. Protocols and applications of snake venomics are reviewed and discussed.

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Venom gland EST analysis of the saw-scaled viper, Echis ocellatus, reveals novel α9β1 integrin-binding motifs in venom metalloproteinases and a new group of putative toxins, renin-like aspartic proteases

Cited by 97 publications

References 52 publications

Medically important differences in snake venom composition are dictated by distinct postgenomic mechanisms

Medically important differences in snake venom composition are dictated by distinct postgenomic mechanisms

Snake venomics of the South and Central American Bushmasters. Comparison of the toxin composition of Lachesis muta gathered from proteomic versus transcriptomic analysis

Snake venomics. Strategy and applications

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