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Combined venom gland cDNA sequencing and venomics of the New Guinea small-eyed snake, Micropechis ikaheka.
J Proteomics. 2014 Oct 14; 110:209-29.JP

Abstract

The venom arsenal of the New Guinea small-eyed snake, Micropechis ikaheka, was investigated by a joint cDNA sequencing and venomics approach. Twenty-seven full-length DNA sequences encoding novel venom proteins were recovered in this study. Using this cDNA dataset we achieved locus-specific resolution for 19 out of the approximately 50 reverse-phase- and SDS-PAGE-separated venom proteins. The venom proteome of M. ikaheka is dominated by at least 29 D49-phospholipase A₂ (PLA₂) and 14 short and long neurotoxins of the three-finger toxin (3FTx) family. These protein classes represent, respectively, 80% and 9.2% of the total venom proteins. Two PIII-metalloproteinase (SVMP) molecules (7.6%), three CRISP isoforms (1.8%), and a single Kunitz-type inhibitor, vespryn, 5'-nucleotidase, serine proteinase and LAO molecules, none of which represents more than 0.7% of the total venom proteome, complete the protein arsenal of M. ikaheka. In concordance with clinical observations, this venom composition points to a central role for post-synaptically-acting neurotoxic toxins in the envenomation strategy developed by this species. PLA₂ molecules represent the main myotoxic components of M. ikaheka venom. In addition, the estimated LD₅₀ for mice of the reverse-phase-isolated 3FTx (0.22 mg/kg) and PLA₂ (1.62 mg/kg) enriched fractions, strongly suggests that these two toxin classes contribute synergistically to venom lethality, with the 3FTxs playing a dominant role. The high structural and functional conservation exhibited by M. ikaheka and Australian elapid venoms may underlay the positive clinical outcomes of envenoming resulting from bites by M. ikaheka that have been documented through the use of bioCSL polyvalent antivenom.

BIOLOGICAL SIGNIFICANCE

The poorly understood venom proteome of the New Guinea small-eyed snake, Micropechis ikaheka, a large and powerfully built elapid endemic to Papua New Guinea and Indonesian West Papua province, was investigated through a combined venomics and venom gland transcriptomics approach. Although M. ikaheka accounts for only a small proportion of snakebites on the mainland, 40% of snakebites on Karkar Island are attributed to bites by this snake. Major effects of envenomings include life-threatening post-synaptic neuromuscular blockade resulting in respiratory paralysis, myotoxicity, severe bleeding, hypotension and cardiovascular abnormalities. We have investigated the contribution of 3FTxs and PLA₂ molecules in venom lethality, myotoxicity, and cardiovascular function. Our work provides important correlations between venom composition and its pharmacological activity. In conjunction with the antivenomics work reported in the companion paper, our study may contribute to improve treatment outcomes for snakebite victims of M. ikaheka.

Authors+Show Affiliations

School of Medicine & Health Sciences, University of Papua New Guinea, Boroko, NCD, Papua New Guinea.Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas (CSIC), Valencia, Spain.Cardiovascular Therapeutics Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia; Australian Venom Research Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia.Cardiovascular Therapeutics Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia; Australian Venom Research Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia.Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas (CSIC), Valencia, Spain.Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.School of Medicine & Health Sciences, University of Papua New Guinea, Boroko, NCD, Papua New Guinea; Australian Venom Research Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia. Electronic address: david.williams@unimelb.edu.au.Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas (CSIC), Valencia, Spain. Electronic address: jcalvete@ibv.csic.es.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25109465

Citation

Paiva, Owen, et al. "Combined Venom Gland cDNA Sequencing and Venomics of the New Guinea Small-eyed Snake, Micropechis Ikaheka." Journal of Proteomics, vol. 110, 2014, pp. 209-29.
Paiva O, Pla D, Wright CE, et al. Combined venom gland cDNA sequencing and venomics of the New Guinea small-eyed snake, Micropechis ikaheka. J Proteomics. 2014;110:209-29.
Paiva, O., Pla, D., Wright, C. E., Beutler, M., Sanz, L., Gutiérrez, J. M., Williams, D. J., & Calvete, J. J. (2014). Combined venom gland cDNA sequencing and venomics of the New Guinea small-eyed snake, Micropechis ikaheka. Journal of Proteomics, 110, 209-29. https://doi.org/10.1016/j.jprot.2014.07.019
Paiva O, et al. Combined Venom Gland cDNA Sequencing and Venomics of the New Guinea Small-eyed Snake, Micropechis Ikaheka. J Proteomics. 2014 Oct 14;110:209-29. PubMed PMID: 25109465.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Combined venom gland cDNA sequencing and venomics of the New Guinea small-eyed snake, Micropechis ikaheka. AU - Paiva,Owen, AU - Pla,Davinia, AU - Wright,Christine E, AU - Beutler,Markus, AU - Sanz,Libia, AU - Gutiérrez,José María, AU - Williams,David J, AU - Calvete,Juan J, Y1 - 2014/08/08/ PY - 2014/06/01/received PY - 2014/07/04/revised PY - 2014/07/14/accepted PY - 2014/8/12/entrez PY - 2014/8/12/pubmed PY - 2015/7/30/medline KW - Micropechis ikaheka KW - Pharmacological effects of snake venom KW - Phospholipase A(2) KW - Snake venomics KW - Three-finger toxin KW - Venom gland cDNA SP - 209 EP - 29 JF - Journal of proteomics JO - J Proteomics VL - 110 N2 - UNLABELLED: The venom arsenal of the New Guinea small-eyed snake, Micropechis ikaheka, was investigated by a joint cDNA sequencing and venomics approach. Twenty-seven full-length DNA sequences encoding novel venom proteins were recovered in this study. Using this cDNA dataset we achieved locus-specific resolution for 19 out of the approximately 50 reverse-phase- and SDS-PAGE-separated venom proteins. The venom proteome of M. ikaheka is dominated by at least 29 D49-phospholipase A₂ (PLA₂) and 14 short and long neurotoxins of the three-finger toxin (3FTx) family. These protein classes represent, respectively, 80% and 9.2% of the total venom proteins. Two PIII-metalloproteinase (SVMP) molecules (7.6%), three CRISP isoforms (1.8%), and a single Kunitz-type inhibitor, vespryn, 5'-nucleotidase, serine proteinase and LAO molecules, none of which represents more than 0.7% of the total venom proteome, complete the protein arsenal of M. ikaheka. In concordance with clinical observations, this venom composition points to a central role for post-synaptically-acting neurotoxic toxins in the envenomation strategy developed by this species. PLA₂ molecules represent the main myotoxic components of M. ikaheka venom. In addition, the estimated LD₅₀ for mice of the reverse-phase-isolated 3FTx (0.22 mg/kg) and PLA₂ (1.62 mg/kg) enriched fractions, strongly suggests that these two toxin classes contribute synergistically to venom lethality, with the 3FTxs playing a dominant role. The high structural and functional conservation exhibited by M. ikaheka and Australian elapid venoms may underlay the positive clinical outcomes of envenoming resulting from bites by M. ikaheka that have been documented through the use of bioCSL polyvalent antivenom. BIOLOGICAL SIGNIFICANCE: The poorly understood venom proteome of the New Guinea small-eyed snake, Micropechis ikaheka, a large and powerfully built elapid endemic to Papua New Guinea and Indonesian West Papua province, was investigated through a combined venomics and venom gland transcriptomics approach. Although M. ikaheka accounts for only a small proportion of snakebites on the mainland, 40% of snakebites on Karkar Island are attributed to bites by this snake. Major effects of envenomings include life-threatening post-synaptic neuromuscular blockade resulting in respiratory paralysis, myotoxicity, severe bleeding, hypotension and cardiovascular abnormalities. We have investigated the contribution of 3FTxs and PLA₂ molecules in venom lethality, myotoxicity, and cardiovascular function. Our work provides important correlations between venom composition and its pharmacological activity. In conjunction with the antivenomics work reported in the companion paper, our study may contribute to improve treatment outcomes for snakebite victims of M. ikaheka. SN - 1876-7737 UR - https://www.unboundmedicine.com/medline/citation/25109465/Combined_venom_gland_cDNA_sequencing_and_venomics_of_the_New_Guinea_small_eyed_snake_Micropechis_ikaheka_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1874-3919(14)00380-7 DB - PRIME DP - Unbound Medicine ER -