1. Venom Composition in a Phenotypically Variable Pit Viper ( Trimeresurus insularis) across the Lesser Sunda Archipelago.
- Author
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Jones BK, Saviola AJ, Reilly SB, Stubbs AL, Arida E, Iskandar DT, McGuire JA, Yates JR 3rd, and Mackessy SP
- Subjects
- Animals, Antivenins pharmacology, Conserved Sequence, Crotalid Venoms isolation & purification, Electrophoresis, Polyacrylamide Gel, Fibrinogen chemistry, Gelatin chemistry, Gene Expression, Indonesia, Islands, L-Amino Acid Oxidase antagonists & inhibitors, L-Amino Acid Oxidase genetics, L-Amino Acid Oxidase metabolism, Lectins, C-Type antagonists & inhibitors, Lectins, C-Type genetics, Lectins, C-Type isolation & purification, Lectins, C-Type metabolism, Membrane Glycoproteins antagonists & inhibitors, Membrane Glycoproteins genetics, Membrane Glycoproteins isolation & purification, Membrane Glycoproteins metabolism, Metalloproteases antagonists & inhibitors, Metalloproteases genetics, Metalloproteases metabolism, Phenotype, Phospholipases A2 genetics, Phospholipases A2 isolation & purification, Phospholipases A2 metabolism, Phosphoric Diester Hydrolases genetics, Phosphoric Diester Hydrolases metabolism, Phylogeny, Proteolysis, Serine Proteases genetics, Serine Proteases metabolism, Trimeresurus genetics, Crotalid Venoms chemistry, L-Amino Acid Oxidase isolation & purification, Metalloproteases isolation & purification, Phosphoric Diester Hydrolases isolation & purification, Serine Proteases isolation & purification, Trimeresurus metabolism
- Abstract
The genus Trimeresurus comprises a group of venomous pitvipers endemic to Southeast Asia and the Pacific Islands. Of these, Trimeresurus insularis, the White-lipped Island Pitviper, is a nocturnal, arboreal species that occurs on nearly every major island of the Lesser Sunda archipelago. In the current study, venom phenotypic characteristics of T. insularis sampled from eight Lesser Sunda Islands (Flores, Lembata, Lombok, Pantar, Sumba, Sumbawa, Timor, and Wetar) were evaluated via SDS-PAGE, enzymatic activity assays, fibrinogenolytic assays, gelatin zymography, and RP-HPLC, and the Sumbawa sample was characterized by venomic analysis. For additional comparative analyses, venoms were also examined from several species in the Trimeresurus complex, including T. borneensis, T. gramineus, T. puniceus, T. purpureomaculatus, T. stejnegeri, and Protobothrops flavoviridis. Despite the geographical isolation, T. insularis venoms from all eight islands demonstrated remarkable similarities in gel electrophoretic profiles and RP-HPLC patterns, and all populations had protein bands in the mass ranges of phosphodiesterases (PDE), l-amino acid oxidases (LAAO), P-III snake venom metalloproteinases (SVMP), serine proteases, cysteine-rich secretory proteins (CRISP), phospholipases A
2 (PLA2 ), and C-type lectins. An exception was observed in the Lombok sample, which lacked protein bands in the mass range of serine protease and CRISP. Venomic analysis of the Sumbawa venom also identified these protein families, in addition to several proteins of lesser abundance (<1%), including glutaminyl cyclase, aminopeptidase, PLA2 inhibitor, phospholipase B, cobra venom factor, 5'-nucleotidase, vascular endothelial growth factor, and hyaluronidase. All T. insularis venoms exhibited similarities in thrombin-like and PDE activities, while significant differences were observed for LAAO, SVMP, and kallikrein-like activities, though these differences were only observed for a few islands. Slight but noticeable differences were also observed with fibrinogen and gelatin digestion activities. Trimeresurus insularis venoms exhibited overall similarity to the other Trimeresurus complex species examined, with the exception of P. flavoviridis venom, which showed the greatest overall differentiation. Western blot analysis revealed that all major T. insularis venom proteins were recognized by Green Pitviper ( T. albolabris) antivenom, and reactivity was also seen with most venom proteins of the other Trimeresurus species, but incomplete antivenom-venom recognition was observed against P. flavoviridis venom proteins. These results demonstrate significant conservation in the venom composition of T. insularis across the Lesser Sunda archipelago relative to the other Trimeresurus species examined.- Published
- 2019
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