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      Variations in neurotoxicity and proteome profile of Malayan krait ( Bungarus candidus) venoms

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          Abstract

          Malayan krait ( Bungarus candidus) is a medically important snake species found in Southeast Asia. The neurotoxic effects of envenoming present as flaccid paralysis of skeletal muscles. It is unclear whether geographical variation in venom composition plays a significant role in the degree of clinical neurotoxicity. In this study, the effects of geographical variation on neurotoxicity and venom composition of B. candidus venoms from Indonesia, Malaysia and Thailand were examined. In the chick biventer cervicis nerve-muscle preparation, all venoms abolished indirect twitches and attenuated contractile responses to nicotinic receptor agonists, with venom from Indonesia displaying the most rapid neurotoxicity. A proteomic analysis indicated that three finger toxins (3FTx), phospholipase A 2 (PLA 2) and Kunitz-type serine protease inhibitors were common toxin groups in the venoms. In addition, venom from Thailand contained L-amino acid oxidase (LAAO), cysteine rich secretory protein (CRISP), thrombin-like enzyme (TLE) and snake venom metalloproteinase (SVMP). Short-chain post-synaptic neurotoxins were not detected in any of the venoms. The largest quantity of long-chain post-synaptic neurotoxins and non-conventional toxins was found in the venom from Thailand. Analysis of PLA 2 activity did not show any correlation between the amount of PLA 2 and the degree of neurotoxicity of the venoms. Our study shows that variation in venom composition is not limited to the degree of neurotoxicity. This investigation provides additional insights into the geographical differences in venom composition and provides information that could be used to improve the management of Malayan krait envenoming in Southeast Asia.

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          Most cited references37

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          Structure, function and evolution of three-finger toxins: mini proteins with multiple targets.

          Snake venoms are complex mixtures of pharmacologically active peptides and proteins. These protein toxins belong to a small number of superfamilies of proteins. Three-finger toxins belong to a superfamily of non-enzymatic proteins found in all families of snakes. They have a common structure of three beta-stranded loops extending from a central core containing all four conserved disulphide bonds. Despite the common scaffold, they bind to different receptors/acceptors and exhibit a wide variety of biological effects. Thus, the structure-function relationships of this group of toxins are complicated and challenging. Studies have shown that the functional sites in these 'sibling' toxins are located on various segments of the molecular surface. Targeting to a wide variety of receptors and ion channels and hence distinct functions in this group of mini proteins is achieved through a combination of accelerated rate of exchange of segments as well as point mutations in exons. In this review, we describe the structural and functional diversity, structure-function relationships and evolution of this group of snake venom toxins. (c) 2010 Elsevier Ltd. All rights reserved.
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            Molecular moulds with multiple missions: functional sites in three-finger toxins.

            R. Kini (2002)
            1. Snake venoms are complex mixtures of pharmacologically active peptides and proteins. 2. These protein toxins belong to a small number of superfamilies of proteins. The present review describes structure-function relationships of three-finger toxins. 3. All toxins share a common structure of three beta-stranded loops extending from a central core. However, they bind to different receptors/acceptors and exhibit a wide variety of biological effects. 4. Thus, the structure-function relationships of this group of toxins are complicated and challenging. 5. Studies have shown that the functional sites in these "sibling" toxins are located on various segments of the molecular surface.
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              Envenoming bites by kraits: the biological basis of treatment-resistant neuromuscular paralysis.

              Beta-bungarotoxin, a neurotoxic phospholipase A2 is a major fraction of the venom of kraits. The toxin was inoculated into one hind limb of young adult rats. The inoculated hind limb was paralysed within 3 h, and remained paralysed for 2 days. The paralysis was associated with the loss of synaptic vesicles from motor nerve terminal boutons, a decline in immunoreactivity of synaptophysin, SNAP-25 and syntaxin, a loss of muscle mass and the upregulation of NaV(1.5) mRNA and protein. Between 3 and 6 h after the inoculation of toxin, some nerve terminal boutons exhibited clear signs of degeneration. Others appeared to be in the process of withdrawing from the synaptic cleft and some boutons were fully enwrapped in terminal Schwann cell processes. By 12 h all muscle fibres were denervated. Re-innervation began at 3 days with the appearance of regenerating nerve terminals, a return of neuromuscular function in some muscles and a progressive increase in the immunoreactivity of synaptophysin, SNAP-25 and syntaxin. Full recovery occurred at 7 days. The data were compared with recently published clinical data on envenoming bites by kraits and by extrapolation we suggest that the acute, reversible denervation caused by beta-bungarotoxin is a credible explanation for the clinically important, profound treatment-resistant neuromuscular paralysis seen in human subjects bitten by these animals.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: ValidationRole: VisualizationRole: Writing – original draft
                Role: ConceptualizationRole: MethodologyRole: ResourcesRole: Supervision
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Software
                Role: Data curationRole: InvestigationRole: Methodology
                Role: MethodologyRole: Resources
                Role: MethodologyRole: Resources
                Role: SupervisionRole: ValidationRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: Writing – original draft
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                30 December 2019
                2019
                : 14
                : 12
                : e0227122
                Affiliations
                [1 ] Kulliyyah of Pharmacy, International Islamic University Malaysia, Kuantan Campus, Bandar Indera Mahkota, Kuantan, Pahang Darul Makmur, Malaysia
                [2 ] Jeffrey Cheah School of Medicine and Health Sciences, Monash University Sunway Campus, Bandar Sunway, Malaysia
                [3 ] Department of Microbiology, Faculty of Science, Mahidol University, Bangkok, Thailand
                [4 ] Snake Farm, Queen Saovabha Memorial Institute, Thai Red Cross Society, Bangkok, Thailand
                [5 ] Monash Venom Group, Department of Pharmacology, Biomedical Discovery Institute, Monash University, Clayton, VIC, Australia
                [6 ] Department of Pharmacology, Phramongkutklao College of Medicine, Bangkok, Thailand
                Instituto Butantan, BRAZIL
                Author notes

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

                Author information
                http://orcid.org/0000-0002-2367-8792
                Article
                PONE-D-19-14681
                10.1371/journal.pone.0227122
                6936869
                31887191
                2f6b6830-2840-4dc6-af51-abb8e239e006
                © 2019 Rusmili 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
                : 24 May 2019
                : 12 December 2019
                Page count
                Figures: 7, Tables: 1, Pages: 19
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100004396, Thailand Research Fund;
                Award ID: TRG6080009
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100010897, Newton Fund;
                Award ID: PDG61W0015
                Award Recipient :
                Funded by: International Islamic University Malaysia
                Award ID: RIGS 17-007-0582
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100000925, National Health and Medical Research Council;
                Award ID: 1110343
                Award Recipient :
                We gratefully acknowledge the following funding: Office of Research Development, Phramongkutklao College of Medicine & Phramongkutklao Hospital (ORD, PCM & PMK, Bangkok, Thailand); Thailand Research Fund (TRG6080009); British Council and Newton Fund for Thailand-UK researcher Link award 2017-18 (PDG61W0015); Research Incentive Grant Scheme of International Islamic University Malaysia (Grant no.: RIGS 17-007-0582); Australian National Health and Medical Research Council (NHMRC) Centres for Research Excellence Grant ID: 1110343. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
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                Biology and Life Sciences
                Toxicology
                Toxic Agents
                Toxins
                Venoms
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Toxicology
                Toxic Agents
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                Biology and Life Sciences
                Toxicology
                Neurotoxicology
                Neurotoxins
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                Pathology and Laboratory Medicine
                Toxicology
                Neurotoxicology
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                Toxicology
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