{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T15:47:47Z","timestamp":1772552867258,"version":"3.50.1"},"reference-count":56,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2017,12,12]],"date-time":"2017-12-12T00:00:00Z","timestamp":1513036800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The spine-bellied sea snake (Hydrophis curtus) is known to cause human deaths, yet its venom composition has not yet been proteomically characterised. An in-depth proteomic analysis was performed on H. curtus venom from two different seasons, January and June, corresponding to adults and subadults, respectively. Venoms from adult and subadult H. curtus individuals were compared using reversed-phase high-performance liquid chromatography (RP-HPLC), matrix-assisted laser desorption ionisation-time of flight (MALDI-TOF) mass spectrometry and liquid chromatography electrospray ionisation mass spectrometry (LC-ESI-MS) to detect intraspecific variation, and the molecular weight data obtained with ESI-MS were used to assess toxin diversity. RP-HPLC and LC-ESI-MS\/MS were used to characterise the venom proteome and estimate the relative abundances of protein families present. The most abundant protein family in January and June venoms is phospholipase A2 (PLA2: January 66.7%; June 54.5%), followed by three-finger toxins (3FTx: January 30.4%; June 40.4%) and a minor component of cysteine-rich secretory proteins (CRISP: January 2.5%; June 5%). Trace amounts of snake venom metalloproteinases (SVMP), C-type lectins and housekeeping and regulatory proteins were also found. Although the complexity of the venom is low by number of families present, each family contained a more diverse set of isoforms than previously reported, a finding that may have implications for the development of next-generation sea snake antivenoms. Intraspecific variability was shown to be minor with one obvious exception of a 14,157-Da protein that was present in some January (adult) venoms, but not at all in June (subadult) venoms. There is also a greater abundance of short-chain neurotoxins in June (subadult) venom compared with January (adult) venom. These differences potentially indicate the presence of seasonal, ontogenetic or sexual variation in H. curtus venom.<\/jats:p>","DOI":"10.3390\/ijms18122695","type":"journal-article","created":{"date-parts":[[2017,12,12]],"date-time":"2017-12-12T13:35:00Z","timestamp":1513085700000},"page":"2695","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["The Venom of the Spine-Bellied Sea Snake (Hydrophis curtus): Proteome, Toxin Diversity and Intraspecific Variation"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0872-2940","authenticated-orcid":false,"given":"Vanessa","family":"Neale","sequence":"first","affiliation":[{"name":"College of Public Health, Medical and Veterinary Sciences, James Cook University, McGregor Road, Smithfield, Cairns 4878, Australia"},{"name":"Australian Institute of Tropical Health and Medicine (AITHM) and Centre for Biodiscovery and Molecular Development of Therapeutics (CBMDT), James Cook University, McGregor Road, Smithfield, Cairns 4878, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1443-7233","authenticated-orcid":false,"given":"Javier","family":"Sotillo","sequence":"additional","affiliation":[{"name":"Australian Institute of Tropical Health and Medicine (AITHM) and Centre for Biodiscovery and Molecular Development of Therapeutics (CBMDT), James Cook University, McGregor Road, Smithfield, Cairns 4878, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jamie E.","family":"Seymour","sequence":"additional","affiliation":[{"name":"Australian Institute of Tropical Health and Medicine (AITHM) and Centre for Biodiscovery and Molecular Development of Therapeutics (CBMDT), James Cook University, McGregor Road, Smithfield, Cairns 4878, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Wilson","sequence":"additional","affiliation":[{"name":"Australian Institute of Tropical Health and Medicine (AITHM) and Centre for Biodiscovery and Molecular Development of Therapeutics (CBMDT), James Cook University, McGregor Road, Smithfield, Cairns 4878, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2017,12,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1016\/j.ympev.2012.09.021","article-title":"Multilocus phylogeny and recent rapid radiation of the viviparous sea snakes (Elapidae: Hydrophiinae)","volume":"66","author":"Sanders","year":"2013","journal-title":"Mol. Phylogenet. Evol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Rasmussen, A.R., Murphy, J.C., Ompi, M., Gibbons, J.W., and Uetz, P. (2011). Marine reptiles. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0027373"},{"key":"ref_3","unstructured":"Meier, J., and White, J. (1995). Clinical toxicology of sea snakebites. Handbook of Clinical Toxicology of Animal Venoms and Poisons, CRC Press."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1093\/icb\/23.2.411","article-title":"Feeding strategies in marine snakes: An analysis of evolutionary, morphological, behavioral and ecological relationships","volume":"23","author":"Voris","year":"1983","journal-title":"Am. Zool."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2047","DOI":"10.1002\/rcm.1148","article-title":"Analysis of Colubroidea snake venoms by liquid chromatography with mass spectrometry: Evolutionary and toxinological implications","volume":"17","author":"Fry","year":"2003","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/j.toxicon.2015.07.008","article-title":"Danger in the reef: Proteome, toxicity, and neutralization of the venom of the olive sea snake, Aipysurus laevis","volume":"107","author":"Laustsen","year":"2015","journal-title":"Toxicon"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"4091","DOI":"10.1016\/j.jprot.2012.05.026","article-title":"Snake venomics of two poorly known Hydrophiinae: Comparative proteomics of the venoms of terrestrial Toxicocalamus longissimus and marine Hydrophis cyanocinctus","volume":"75","author":"Calvete","year":"2012","journal-title":"J. Proteom."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.jprot.2014.03.034","article-title":"Two color morphs of the pelagic yellow-bellied sea snake, Pelamis platura, from different locations of Costa Rica: Snake venomics, toxicity, and neutralization by antivenom","volume":"103","author":"Lomonte","year":"2014","journal-title":"J. Proteom."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.jprot.2015.05.035","article-title":"Venomics of the beaked sea snake, Hydrophis schistosus: A minimalist toxin arsenal and its cross-neutralization by heterologous antivenoms","volume":"126","author":"Tan","year":"2015","journal-title":"J. Proteom."},{"key":"ref_10","unstructured":"Dunson, W.A. (1975). Adaptations of sea snakes. Biology of Sea Snakes, University Park Press."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1284","DOI":"10.1136\/bmj.1.5235.1284","article-title":"Myoglobinuria and sea-snake-bite poisoning","volume":"1","author":"Reid","year":"1961","journal-title":"Br. Med. J."},{"key":"ref_12","unstructured":"Dunson, W.A. (1975). Epidemiology and clinical aspects of sea snake bites. Biology of Sea Snakes, University Park Press."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1021\/jf60191a034","article-title":"Sea snake venoms and neurotoxins","volume":"22","author":"Tu","year":"1974","journal-title":"J. Agric. Food Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1111\/zsc.12070","article-title":"Multilocus phylogeography of the sea snake Hydrophis curtus reveals historical vicariance and cryptic lineage diversity","volume":"43","author":"Ukuwela","year":"2014","journal-title":"Zool. Scr."},{"key":"ref_15","unstructured":"Gopalakrishnakone, P. (1994). Sea snake bites in the Asia-Pacific region. Sea Snake Toxinology, Singapore University Press."},{"key":"ref_16","unstructured":"Faiz, A., Fernando, R., Gnanathasan, C.A., Habib, A.G., and Yang, C.-C. (2015). Venomous snakes and snakebites in India. Clinical Toxinology in Asia Pacific and Africa, Springer Science + Business Media."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Cogger, H.G. (2014). Reptiles and Amphibians of Australia, CSIRO Publishing. [7th ed.].","DOI":"10.1071\/9780643109773"},{"key":"ref_18","unstructured":"Dunson, W.A. (1975). Attacks by sea snakes on divers. Biology of Sea Snakes, University Park Press."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1016\/j.toxlet.2011.05.1028","article-title":"The study on venom proteins of Lapemis hardwickii by cDNA phage display","volume":"206","author":"Tan","year":"2011","journal-title":"Toxicol. Lett."},{"key":"ref_20","first-page":"345","article-title":"Diversity of PLA2 genes from sea snake Lapemis hardwickii Gray venom","volume":"33","author":"Yang","year":"2001","journal-title":"Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1016\/0003-9861(79)90047-X","article-title":"Conformational analysis of a snake neurotoxin by prediction from sequence, circular dichroism, and Raman spectroscopy","volume":"193","author":"Fox","year":"1979","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_22","first-page":"457","article-title":"Identification and functional characterization of three postsynaptic short-chain neurotoxins from Hydrophiinae, Lapemis hardwickii Gray","volume":"33","author":"Zhong","year":"2001","journal-title":"Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao"},{"key":"ref_23","first-page":"5","article-title":"Cloning and sequence analysis of full-length cDNA encoding cysteine-rich venom protein from Lapemis hardwickii","volume":"40","author":"Wei","year":"2001","journal-title":"Acta Sci. Nat. Univ. Sunyatseni"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2965","DOI":"10.1016\/j.febslet.2007.05.047","article-title":"Identification and characterization of Harobin, a novel fibrino(geno)lytic serine protease from a sea snake (Lapemis hardwickii)","volume":"581","author":"He","year":"2007","journal-title":"FEBS Lett."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/j.toxicon.2004.05.022","article-title":"The in vitro neuromuscular activity of Indo-Pacific sea-snake venoms: Efficacy of two commercially available antivenoms","volume":"44","author":"Chetty","year":"2004","journal-title":"Toxicon"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"307","DOI":"10.3923\/jpt.2007.307.318","article-title":"Histopathological changes induced in mice after intramuscular and intraperitoneal injections of venom from spine-bellied sea snake, Lapemis curtus (Shaw, 1802)","volume":"2","author":"Karthikeyan","year":"2007","journal-title":"J. Pharmacol. Toxicol."},{"key":"ref_27","first-page":"351","article-title":"A comparative study of the biological properties of some sea snake venoms","volume":"99","author":"Tan","year":"1991","journal-title":"Comp. Biochem. Physiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1186\/1471-2148-7-175","article-title":"Expression pattern of three-finger toxin and phospholipase A2 genes in the venom glands of two sea snakes, Lapemis curtus and Acalyptophis peronii: Comparison of evolution of these toxins in land snakes, sea kraits and sea snakes","volume":"7","author":"Pahari","year":"2007","journal-title":"BMC Evol. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1586\/epr.11.61","article-title":"Proteomic tools against the neglected pathology of snake bite envenoming","volume":"8","author":"Calvete","year":"2011","journal-title":"Expert Rev. Proteom."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1279","DOI":"10.1016\/0041-0101(91)90116-9","article-title":"Snake venom variability: Methods of study, results and interpretation","volume":"29","author":"Chippaux","year":"1991","journal-title":"Toxicon"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3556","DOI":"10.1021\/pr800332p","article-title":"Snake venomics of the lancehead pitviper Bothrops asper: Geographic, individual, and ontogenetic variations","volume":"7","author":"Sanz","year":"2008","journal-title":"J. Proteome Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/0041-0101(91)90106-2","article-title":"Comparative study of nine Bothrops snake venoms from adult female snakes and their offspring","volume":"29","author":"Furtado","year":"1991","journal-title":"Toxicon"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"92","DOI":"10.2307\/1445927","article-title":"Venom ontogeny in the Pacific rattlesnakes Crotalus viridis helleri and C. v. oreganus","volume":"1988","author":"Mackessy","year":"1988","journal-title":"Copeia"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4218","DOI":"10.1002\/pmic.201100287","article-title":"Bothrops jararaca venom proteome rearrangement upon neonate to adult transition","volume":"11","author":"Zelanis","year":"2011","journal-title":"Proteomics"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1016\/j.jprot.2014.02.020","article-title":"Venomous snakes of Costa Rica: Biological and medical implications of their venom proteomic profiles analyzed through the strategy of snake venomics","volume":"105","author":"Lomonte","year":"2014","journal-title":"J. Proteom."},{"key":"ref_36","unstructured":"World Health Organization (2010). WHO Guidelines for the Production, Control and Regulation of Snake Antivenom Immunoglobulins, World Health Organization."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1016\/j.toxicon.2005.11.007","article-title":"Sex-based individual variation of snake venom proteome among eighteen Bothrops jararaca siblings","volume":"47","author":"Menezes","year":"2006","journal-title":"Toxicon"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1034","DOI":"10.1002\/rcm.2931","article-title":"Mass spectrometric analysis of the individual variability of Bothrops jararaca venom peptide fraction. Evidence for sex-based variation among the bradykinin-potentiating peptides","volume":"21","author":"Pimenta","year":"2007","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1265","DOI":"10.1074\/mcp.M500061-MCP200","article-title":"Exponentially modified protein abundance index (emPAI) for estimation of absolute protein amount in proteomics by the number of sequenced peptides per protein","volume":"4","author":"Ishihama","year":"2005","journal-title":"Mol. Cell. Proteom."},{"key":"ref_40","unstructured":"Possani, L.D., Schwartz, E.F., and Rodr\u00edguez de la Vega, R.C. (2015). Modern venom profiling: Mining into scorpion venom biodiversity. Scorpion Venoms, Springer."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.toxicon.2013.01.019","article-title":"Alpha neurotoxins","volume":"66","author":"Barber","year":"2013","journal-title":"Toxicon"},{"key":"ref_42","unstructured":"White, J. (2013). A Clinician\u2019s Guide to Australian Venomous Bites and Stings: Incorporating the Updated CSL Antivenom Handbook, bioCSL."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"745","DOI":"10.1016\/0035-9203(89)90313-1","article-title":"Snake venoms in science and clinical medicine 3. Neuropharmacological aspects of the activity of snake venoms","volume":"83","author":"Harris","year":"1989","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"571","DOI":"10.1113\/expphysiol.1987.sp003098","article-title":"Effects of phospholipase of Enhydrina schistosa venom on nerve, motor end-plate and muscle of the mouse","volume":"72","author":"Brook","year":"1987","journal-title":"Q. J. Exp. Physiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1016\/0041-0101(78)90191-5","article-title":"Ultrastructural changes in skeletal muscle caused by a phospholipase A2 fraction isolated from the venom of a sea snake, Enhydrina schistosa","volume":"16","author":"Geh","year":"1978","journal-title":"Toxicon"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2897","DOI":"10.1007\/s00018-008-8113-3","article-title":"Cellular pathology induced by snake venom phospholipase A2 myotoxins and neurotoxins: Common aspects of their mechanisms of action","volume":"65","author":"Montecucco","year":"2008","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1111\/j.1432-1033.1980.tb04982.x","article-title":"Correlation of enzymatic activity and anticoagulant properties of phospholipase A2","volume":"112","author":"Verheij","year":"1980","journal-title":"Eur. J. Biochem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/j.biochi.2009.12.006","article-title":"Isolation of an acidic phospholipase A2 from the venom of the snake Bothrops asper of Costa Rica: Biochemical and toxicological characterization","volume":"92","author":"Angulo","year":"2010","journal-title":"Biochimie"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Jackson, T., Koludarov, I., Ali, S., Dobson, J., Zdenek, C., Dashevsky, D., op den Brouw, B., Masci, P., Nouwens, A., and Josh, P. (2016). Rapid radiations and the race to redundancy: An investigation of the evolution of Australian elapid snake venoms. Toxins, 8.","DOI":"10.3390\/toxins8110309"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"119","DOI":"10.3724\/SP.J.1245.2014.00119","article-title":"Age-related variation in snake venom: Evidence from two snakes (Naja atra and Deinagkistrodon acutus) in Southeastern China","volume":"5","author":"He","year":"2014","journal-title":"Asian Herpetol. Res."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.toxicon.2016.04.049","article-title":"An analysis of venom ontogeny and prey-specific toxicity in the monocled cobra (Naja kaouthia)","volume":"119","author":"Modahl","year":"2016","journal-title":"Toxicon"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1266","DOI":"10.1021\/pr101040f","article-title":"Snake venomics of African spitting cobras: Toxin composition and assessment of congeneric cross-reactivity of the pan-African EchiTAb-Plus-ICP antivenom by antivenomics and neutralization approaches","volume":"10","author":"Petras","year":"2011","journal-title":"J. Proteome Res."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"2128","DOI":"10.1016\/j.jprot.2012.01.006","article-title":"Comparative proteomic analysis of the venom of the taipan snake, Oxyuranus scutellatus, from Papua New Guinea and Australia: Role of neurotoxic and procoagulant effects in venom toxicity","volume":"75","author":"Herrera","year":"2012","journal-title":"J. Proteom."},{"key":"ref_54","unstructured":"(2017, March 09). NCBI Non-Redundant Serpentes Database, Available online: https:\/\/www.ncbi.nlm.nih.gov\/protein\/?term=serpentes."},{"key":"ref_55","unstructured":"(2017, March 09). UniProtKB Serpentes Database. Available online: http:\/\/www.uniprot.org\/uniprot\/?query=Serpentes&sort=score."},{"key":"ref_56","unstructured":"(2017, November 03). UniProtKB Database. search term \u2018Lapemis hardwickii\u2019."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/12\/2695\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:53:43Z","timestamp":1760208823000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/12\/2695"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,12,12]]},"references-count":56,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2017,12]]}},"alternative-id":["ijms18122695"],"URL":"https:\/\/doi.org\/10.3390\/ijms18122695","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,12,12]]}}}