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      Proteomics and immunocharacterization of Asian mountain pit viper ( Ovophis monticola) venom

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          Abstract

          The venomic profile of Asian mountain pit viper Ovophis monticola is clarified in the present study. Using mass spectrometry-based proteomics, 247 different proteins were identified in crude venom of O. monticola found in Thailand. The most abundant proteins were snake venom metalloproteases (SVMP) (36.8%), snake venom serine proteases (SVSP) (31.1%), and phospholipases A 2 (PLA 2) (12.1%). Less abundant proteins included L-amino acid oxidase (LAAO) (5.7%), venom nerve growth factor (3.6%), nucleic acid degrading enzymes (3.2%), C-type lectins (CTL) (1.6%), cysteine-rich secretory proteins (CRISP) (1.2%) and disintegrin (1.2%). The immunoreactivity of this viper’s venom to a monovalent antivenom against green pit viper Trimeresurus albolabris, or to a polyvalent antivenom against hemotoxic venom was investigated by indirect ELISA and two-dimensional (2D) immunoblotting. Polyvalent antivenom showed substantially greater reactivity levels than monovalent antivenom. A titer for the monovalent antivenom was over 1:1.28x10 7 dilution while that of polyvalent antivenom was 1:5.12x10 7. Of a total of 89 spots comprising 173 proteins, 40 spots of predominantly SVMP, SVSP and PLA 2 were specific antigens for antivenoms. The 49 unrecognized spots containing 72 proteins were characterized as non-reactive proteins, and included certain types of CTLs and CRISPs. These neglected venom constituents could limit the effectiveness of antivenom-based therapy currently available for victims of pit viper envenomation.

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          Most cited references47

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          Enzymatic toxins from snake venom: structural characterization and mechanism of catalysis.

          Snake venoms are cocktails of enzymes and non-enzymatic proteins used for both the immobilization and digestion of prey. The most common snake venom enzymes include acetylcholinesterases, l-amino acid oxidases, serine proteinases, metalloproteinases and phospholipases A(2) . Higher catalytic efficiency, thermal stability and resistance to proteolysis make these enzymes attractive models for biochemists, enzymologists and structural biologists. Here, we review the structures of these enzymes and describe their structure-based mechanisms of catalysis and inhibition. Some of the enzymes exist as protein complexes in the venom. Thus we also discuss the functional role of non-enzymatic subunits and the pharmacological effects of such protein complexes. The structures of inhibitor-enzyme complexes provide ideal platforms for the design of potent inhibitors which are useful in the development of prototypes and lead compounds with potential therapeutic applications. © 2011 The Authors Journal compilation © 2011 FEBS.
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            Snakebite envenoming

            This corrects the article DOI: 10.1038/nrdp.2017.63.
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              Venom of the Brown Treesnake, Boiga irregularis: ontogenetic shifts and taxa-specific toxicity.

              The Brown Treesnake (Boiga irregularis), a rear-fanged member of the polyphyletic family Colubridae, is an introduced predator on Guam which has been responsible for numerous human envenomations. Because little is known about this species' venom, we characterized venom proteins from B. irregularis using enzyme assays, one and 2D electrophoresis, Western blot analysis, mass spectrometry, HPLC and toxicity assays. Venom yields and protein content varied significantly with snake size, and large adult specimens averaged over 500 microl venom (19.2 mg, protein content approximately 90%). Only two enzymes, azocaseinolytic metalloprotease and acetylcholinesterase, were detected in venoms, and both activities increased with snake size/age. Western blot analysis demonstrated a 25 kDa CRiSP homolog in venoms from both neonate and adult snakes. 2D electrophoresis showed variation between venoms from neonate and adult snakes, especially with respect to metalloprotease and acetylcholinesterase. Analysis by MALDI-TOF mass spectrometry revealed the presence of numerous proteins with molecular masses of approximately 8.5-11 kDa. Adult B. irregularis venom was quite toxic to domestic chickens (Gallus domesticus; 1.75 microg/g) and lizards (Hemidactylus geckos: 2.5 microg/g and Carlia skinks: 4.5 microg/g), and intoxication was characterized by rapid paralysis of all species and neck droop in chickens. Toxicity of venom from neonates toward geckos was 1.1 microg/g, consistent with the presence of a greater diversity of 8-11 kDa proteins (suspected neurotoxins) in these venoms. All of these values were notably lower than murine LD50 values (neonate: 18 microg/g; adult: 31 microg/g). Like venoms of several front-fanged species, B. irregularis venom showed an ontogenetic shift in enzyme activities and toxicity, and neonate snakes produced more toxic venoms with lower protease and acetylcholinesterase activities. High toxicity toward non-mammalian prey demonstrated the presence of taxa-specific effects (and thus toxins) in B. irregularis venom, likely a characteristic of many colubrid snake venoms. We hypothesize that the lack of significant envenomation effects in humans following most colubrid bites results from this taxa-specific action of colubrid venom components, not from a lack of toxins.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Project administrationRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: MethodologyRole: ResourcesRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: SoftwareRole: ValidationRole: Writing – original draft
                Role: InvestigationRole: Methodology
                Role: ConceptualizationRole: Resources
                Role: ConceptualizationRole: InvestigationRole: Methodology
                Role: InvestigationRole: Methodology
                Role: Resources
                Role: MethodologyRole: Software
                Role: Supervision
                Role: ConceptualizationRole: Formal analysisRole: InvestigationRole: MethodologyRole: SupervisionRole: ValidationRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                1 December 2021
                2021
                : 16
                : 12
                : e0260496
                Affiliations
                [1 ] Department of Biology, Faculty of Science, Mahidol University, Ratchathewi, Bangkok, Thailand
                [2 ] Snake Farm, Queen Saovabha Memorial Institute, The Thai Red Cross Society, Bangkok, Thailand
                [3 ] Department of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol University, Ratchathewi, Bangkok, Thailand
                [4 ] Department of Research and Development, Queen Saovabha Memorial Institute, The Thai Red Cross Society, Bangkok, Thailand
                Universidade Federal de Pernambuco, BRAZIL
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                https://orcid.org/0000-0002-8248-2300
                Article
                PONE-D-21-18686
                10.1371/journal.pone.0260496
                8635378
                34851989
                5b0af93b-66e4-4185-98d4-7979781822b4
                © 2021 Sitprija et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 7 June 2021
                : 10 November 2021
                Page count
                Figures: 4, Tables: 4, Pages: 21
                Funding
                Funded by: Center of Excellence on Biodiversity
                Award ID: BDC-PG4-161009.
                Award Recipient :
                This research was funded by the Center of Excellence on Biodiversity, grant number BDC-PG4-161009.
                Categories
                Research Article
                Biology and Life Sciences
                Toxicology
                Toxic Agents
                Toxins
                Venoms
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Toxicology
                Toxic Agents
                Toxins
                Venoms
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Vertebrates
                Amniotes
                Reptiles
                Squamates
                Snakes
                Biology and Life Sciences
                Zoology
                Animals
                Vertebrates
                Amniotes
                Reptiles
                Squamates
                Snakes
                Biology and Life Sciences
                Biochemistry
                Enzymology
                Enzymes
                Proteases
                Serine Proteases
                Biology and Life Sciences
                Biochemistry
                Proteins
                Enzymes
                Proteases
                Serine Proteases
                Physical Sciences
                Chemistry
                Chemical Compounds
                Organic Compounds
                Amino Acids
                Hydroxyl Amino Acids
                Serine
                Physical Sciences
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                Amino Acids
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                Serine
                Biology and Life Sciences
                Biochemistry
                Proteins
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                Serine
                Biology and Life Sciences
                Biochemistry
                Enzymology
                Enzymes
                Hydrolases
                Esterases
                Phospholipases
                Biology and Life Sciences
                Biochemistry
                Proteins
                Enzymes
                Hydrolases
                Esterases
                Phospholipases
                Biology and Life Sciences
                Biochemistry
                Enzymology
                Enzymes
                Proteases
                Biology and Life Sciences
                Biochemistry
                Proteins
                Enzymes
                Proteases
                Biology and Life Sciences
                Organisms
                Eukaryota
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                Vertebrates
                Amniotes
                Reptiles
                Squamates
                Snakes
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                Biology and Life Sciences
                Zoology
                Animals
                Vertebrates
                Amniotes
                Reptiles
                Squamates
                Snakes
                Vipers
                Biology and Life Sciences
                Biochemistry
                Enzymology
                Enzymes
                Proteases
                Metalloproteases
                Biology and Life Sciences
                Biochemistry
                Proteins
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