title: Proteomics and antivenomics of Papuan black snake (Pseudechis papuanus) venom with analysis of its toxicological profile and the preclinical efficacy of Australian antivenoms.
abstract: The Papuan black snake (Pseudechis papuanus Serpentes: Elapidae) is endemic to Papua New Guinea, Indonesian Papua and Australia's Torres Strait Islands. We have investigated the biological activity and proteomic composition of its venom. The P. papuanus venom proteome is dominated by a variety (n≥18) of PLA2s, which together account for ~90% of the venom proteins, and a set of low relative abundance proteins, including a short-neurotoxic 3FTx (3.1%), 3-4 PIII-SVMPs (2.8%), 3 cysteine-rich secretory proteins (CRISP; 2.3%) 1-3 l-amino acid oxidase (LAAO) molecules (1.6%). Probing of a P. papuanus cDNA library with specific primers resulted in the elucidation of the full-length nucleotide sequences of six new toxins, including vespryn and NGF not found in the venom proteome, and a calglandulin protein involved in toxin expression with the venom glands. Intravenous injection of P. papuanus venom in mice induced lethality, intravascular haemolysis, pulmonary congestion and oedema, and anticoagulation after intravenous injection, and these effects are mainly due to the action of PLA2s. This study also evaluated the in vivo preclinical efficacy of Australian black snake and polyvalent Seqirus antivenoms. These antivenoms were effective in neutralising the lethal, PLA2and anticoagulant activities of P. papuanus venom in mice. On the other hand, all of the Seqirus antivenoms tested using an antivenomic approach exhibited strong immunorecognition of all the venom components. These preclinical results suggest that Australian Seqirus1antivenoms may provide paraspecific protection against P. papuanus venom in humans.
  
  
  
  The toxicological profile and proteomic composition of the venom of the Papuan black snake, Pseudechis papuanus, a large diurnal snake endemic to the southern coast of New Guinea and a handful of close offshore islands, were investigated. Intravenous injection of P. papuanus venom in mice induced intravascular hemolysis, pulmonary congestion and edema, anticoagulation, and death. These activities could be assigned to the set of PLA2molecules, which dominate the P. papuanus venom proteome. This study also showed that Australian Seqirus black snake or polyvalent antivenoms were effective in neutralising the lethal, PLA2and anticoagulant activities of the venom. These preclinical results support the continued recommendation of these Seqirus antivenoms in the clinical management of P. papuanus envenoming in Australia, Papua New Guinea or Indonesian Papua Province.
authors.affiliation: Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas (CSIC), Valencia, Spain.
authors.forename: Davinia
authors.initials: D
authors.lastname: Pla
authors.affiliation: Charles Campbell Toxinology Centre, School of Medicine & Health Sciences, University of Papua New Guinea, Boroko, NCD, Papua New Guinea.
authors.forename: Benjamin W
authors.initials: BW
authors.lastname: Bande
authors.affiliation: Australian Venom Research Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Ronelle E
authors.initials: RE
authors.lastname: Welton
authors.affiliation: Charles Campbell Toxinology Centre, School of Medicine & Health Sciences, University of Papua New Guinea, Boroko, NCD, Papua New Guinea.
authors.forename: Owen K
authors.initials: OK
authors.lastname: Paiva
authors.affiliation: Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas (CSIC), Valencia, Spain.
authors.forename: Libia
authors.initials: L
authors.lastname: Sanz
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
authors.forename: Álvaro
authors.initials: Á
authors.lastname: Segura
authors.affiliation: Australian Venom Research Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Parkville, Victoria 3010, Australia; Cardiovascular Therapeutics Unit, Department of Pharmacology and Therapeutics, University of Melbourne, Victoria 3010, Australia.
authors.forename: Christine E
authors.initials: CE
authors.lastname: Wright
authors.affiliation: Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas (CSIC), Valencia, Spain. Electronic address: jcalvete@ibv.csic.es.
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Calvete
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica. Electronic address: jose.gutierrez@ucr.ac.cr.
authors.forename: José María
authors.initials: JM
authors.lastname: Gutiérrez
authors.affiliation: Charles Campbell Toxinology Centre, 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, Parkville, Victoria 3010, Australia. Electronic address: david.williams@unimelb.edu.au.
authors.forename: David J
authors.initials: DJ
authors.lastname: Williams
mesh_terms: D000818:Animals; D000997:Antivenins / Q000032:analysis / Q000378:metabolism / Q000627:therapeutic use; D001315:Australia; D004353:Drug Evaluation, Preclinical; D004546:Elapid Venoms / Q000032:analysis* / Q000378:metabolism; D017815:Elapidae / Q000378:metabolism*; D007928:Lethal Dose 50; D051379:Mice; D009500:Neutralization Tests; D010219:Papua New Guinea; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics; D018675:Toxicity Tests; D016896:Treatment Outcome
chemical_list: D000997:Antivenins; D004546:Elapid Venoms; D020543:Proteome
keywords: Antivenomics; In vivo neutralisation; Lethality; Myotoxicity; Pseudechis papuanus; Seqirus antivenoms; Snake venomics