Group IB phospholipase A2 from Pseudonaja textilis

Armugam, Arunmozhiarasi; Gong, Nanling; Li, Xiaojie; Siew, Phui Yee; Chai, Siaw Ching; Nair, Ramkishen; Jeyaseelan, Kandiah

doi:10.1016/j.abb.2003.09.045

Cited by 22 publications

(20 citation statements)

References 50 publications

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“…These are suggestive of post-translational modifications of individual proteins. The abundance of spots in the molecular mass range 10 -15 kDa with low pI corresponds to that expected of PLA 2 s, which have already been shown to be present in P. textilis venom (6,8). The overall pattern of staining was similar when colloidal Coomassie Blue staining was used; however, the relative intensity of some protein spots was different compared with detection by silver staining.…”

Section: Separation and Comparative Analysis Of P Textilis Venommentioning

confidence: 80%

“…The multiple species observed for other proteins is unlikely to be explained by such modifications. For example in the case of PLA 2 s, it is more likely to be due to multiple isoforms for which there is evidence in P. textilis (7,8). Furthermore in the Cy dye overlay experiments there seemed to be a high degree of concordance between treated and untreated samples for the PLA 2 s.…”

Section: Separation and Comparative Analysis Of P Textilis Venommentioning

confidence: 97%

“…These include postsynaptic ␣-neurotoxins (2-4), the presynaptic neurotoxin textilotoxin (5-7), phospholipase A 2 s (PLA 2 s) 1 (6,8); serine protease inhibitors (9,10), and a prothrombin activator (11,12). Fry (13) has written a comprehensive review on the properties of venom components from Australian elapids.…”

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confidence: 99%

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Molecular Diversity in Venom from the Australian Brown Snake, Pseudonaja textilis

Birrell

Earl

Masci

et al. 2006

Molecular & Cellular Proteomics

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Venom from the Australian elapid Pseudonaja textilis (Common or Eastern Brown snake), is the second most toxic snake venom known and is the most common cause of death from snake bite in Australia. This venom is known to contain a prothrombin activator complex, serine proteinase inhibitors, various phospholipase A 2 s, and preand postsynaptic neurotoxins. In this study, we performed a proteomic identification of the venom using two-dimensional gel electrophoresis, mass spectrometry, and de novo peptide sequencing. We identified most of the venom proteins including proteins previously not known to be present in the venom. In addition, we used immunoblotting and post-translational modification-specific enzyme stains and antibodies that reveal the complexity and regional diversity of the venom. Modifications observed include phosphorylation, ␥-carboxylation, and glycosylation. Snakes from the Australian elapid genus Pseudonaja are fast moving and highly venomous and are responsible for most deaths by snake bite in Australia. Of the eight classified species of Pseudonaja, Pseudonaja textilis, the Common or Eastern Brown snake, has the most lethal venom, surpassed only by Oxyuranus microlepidotus, the Inland Taipan. The venom is strongly procoagulant and has little hemolytic or myolytic activity (1). Several researchers have examined the venom from P. textilis and have isolated numerous toxins.These include postsynaptic ␣-neurotoxins (2-4), the presynaptic neurotoxin textilotoxin (5-7), phospholipase A 2 s (PLA 2 s) 1 (6, 8); serine protease inhibitors (9, 10), and a prothrombin activator (11, 12). Fry (13) has written a comprehensive review on the properties of venom components from Australian elapids.Barnett et al.(2) isolated a postsynaptic ␣-neurotoxin from the venom and named it pseudonajatoxin A. This protein is 117 amino acids in length, which is considerably larger than other snake neurotoxins, and like other ␣-neurotoxins acts by binding to acetylcholine receptors. Pseudonajatoxin B by contrast (3) contains only 71 amino acids and has considerable homology with other postsynaptic long ␣-neurotoxins. Six other ␣-neurotoxins have been cloned and expressed from venom gland RNA (4). These have fewer amino acids (57 or 58 residues) and possess lower neurotoxic activities than the short-chain ␣-neurotoxins found in other snakes. Textilotoxin is the most potent neurotoxin yet isolated from the venom of a land snake. It represents 3% by weight of the crude venom and 70% of its total lethality (1). It is comprised of five PLA 2 subunits, two of which are identical, and acts by blocking the release of acetylcholine following the arrival of an action potential at a nerve terminal (14). Armugam et al. (8) isolated four other PLA 2 s from the venom and found them to be group 1B PLA 2 s. In that study, analysis of DNA from the venom gland showed that only two genes and two cDNAs were responsible for the four PLA 2 proteins produced. This is similar to the short-chain ␣-neurotoxins (4) where alternative splicing of the P....

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Section: Separation and Comparative Analysis Of P Textilis Venommentioning

confidence: 80%

Section: Separation and Comparative Analysis Of P Textilis Venommentioning

confidence: 97%

See 1 more Smart Citation

Molecular Diversity in Venom from the Australian Brown Snake, Pseudonaja textilis

Birrell

Earl

Masci

et al. 2006

Molecular & Cellular Proteomics

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“…Advances in transcriptomic and proteomic technologies have contributed to the identification of some of the less abundant toxins within the venom of Australian snakes [19,20]. However, to date only a handful of full-length genomic sequences have been identified for Australian elapids toxins, most notably for the phospholipase A 2 (PLA 2 ) and neurotoxin families from P. textilis [21][22][23]. This study describes the cloning and comparative analysis of two types of kunitz inhibitor along with waprin toxin sequences from a total of eleven Australian elapid snakes, demonstrating a common evolutionary relationship between the three types of toxin.…”

Section: Introductionmentioning

confidence: 99%

Common evolution of waprin and kunitz-like toxin families in Australian venomous snakes

Pierre

Earl

Filippovich

et al. 2008

Cell. Mol. Life Sci.

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The venoms of Australian snakes contain a myriad of pharmacologically active toxin components. This study describes the identification and comparative analysis of two distinct toxin families, the kunitztype serine protease inhibitors and waprins, and demonstrates a previously unknown evolutionary link between the two. Multiple cDNA and full-length gene isoforms were cloned and shown to be composed of three exons separated by two introns. A high degree of identity was observed solely within the first exon which coded for the propeptide sequence and its cleavage site, and indicates that each toxin family has arisen from a gene duplication event followed by diversification only within the portion of the gene coding for the functional toxin. It is proposed that while the mechanism of toxin secretion is highly conserved, diversification of mature toxin sequences allows for the existence of multiple protein isoforms in the venom to adapt to variations within the prey environment.

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“…They are sub-grouped into group IA or group IB depending on the presence of a characteristic elapid or pancreatic loop respectively; the elapid loop form contains a five amino acid deletion [55]. Generally, most elapid PLA 2 belong to group IA though some PLA 2 toxins from Oxyuranus microlepidotus, Pseudonaja textilis and Notechis scutatus have been classified as group IB PLA 2 [56].…”

Section: Pla 2 Toxinsmentioning

confidence: 99%

Deep venomics of the Pseudonaja genus reveals inter- and intra-specific variation

Reeks

Lavergne

Sunagar

et al. 2016

Journal of Proteomics

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Australian elapid venom remains an under-investigated resource of novel bioactive peptides. In this study, the venom gland transcriptomes and proteomes of the Australian western brown snakes, Pseudonaja aspidorhyncha and Pseudonaja nuchalis, were compared to Pseudonaja textilis. A deep venomics strategy incorporating high throughput 454 pyrosequencing gave a total of 200,911 raw reads for the three venoms. Subsequent annotation identified 5716 transcripts from 20 different toxin families with inter-specific variation between species observed in eight of the less abundant families. Integration of each venom proteome with the corresponding annotated reads identified 65 isoforms from six toxin families; high sequence coverage highlighted subtle differences between sequences and intra and inter-specific variation between species. High quality MS/MS data identified unusual glycoforms with natriuretic peptides from P. aspidorhyncha and P. nuchalis containing O-linked trisaccharides with high homology to the glycosylated region of TNPc. Molecular evolutionary assessments indicated the accelerated evolution of all toxin families with the exception of both natriuretic peptides and P. aspidorhyncha PLA2s that were found to be evolutionarily constrained under purifying selection pressures. This study has revealed a wide range of novel peptide sequences from six bioactive peptide families and highlights the subtle differences between toxins in these closely related species. Biological SignificanceMining Australia's vastly untapped source of toxins from its venomous creatures has been significantly advanced by employing deep venomics methodology.Technological advances in transcriptome analysis using next generation sequencing platforms and proteome analysis by highly sensitive tandem mass spectrometry allowed a more comprehensive interrogation of three underinvestigated brown snake (Pseudonaja) venoms uncovering many novel peptide sequences that are unique to these closely related species. This generic strategy will provide invaluable information when applied to other venomous snakes for a deeper understanding of venom composition, envenomation, venom evolution, as well as identifying research tools and drug leads.

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Group IB phospholipase A2 from Pseudonaja textilis

Cited by 22 publications

References 50 publications

Molecular Diversity in Venom from the Australian Brown Snake, Pseudonaja textilis

Molecular Diversity in Venom from the Australian Brown Snake, Pseudonaja textilis

Common evolution of waprin and kunitz-like toxin families in Australian venomous snakes

Deep venomics of the Pseudonaja genus reveals inter- and intra-specific variation

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