Characterization of toxins from the broad-banded water snake Helicops angulatus (Linnaeus, 1758): isolation of a cysteine-rich secretory protein, Helicopsin
| Autor: | Elda E. Sánchez, W. Andy Tao, Jacob A. Galan, Alexis Rodríguez-Acosta, Amalid Estrella, Luis F. Navarrete, Belsy Guerrero |
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| Rok vydání: | 2010 |
| Předmět: |
Health
Toxicology and Mutagenesis Injections Subcutaneous Molecular Sequence Data Neurotoxins Toxicology Peptide Mapping Homology (biology) Salivary Glands Lethal Dose 50 Mice Cysteine-rich secretory protein Tandem Mass Spectrometry Neurotoxin Animals Amino Acid Sequence Cysteine Chromatography High Pressure Liquid Gel electrophoresis Helicops angulatus Mice Inbred BALB C biology Behavior Animal Lethal dose Colubridae General Medicine biology.organism_classification Secretory protein Biochemistry biology.protein Chromatography Gel Nervous System Diseases Colubroidea Snake Venoms |
| Zdroj: | Archives of toxicology. 85(4) |
| ISSN: | 1432-0738 |
| Popis: | Helicops angulatus (broad-banded water snake) according to recent proposals is presently cited in the family Dipsadidae, subfamily Xenodontinae, forming the tribe Hydropsini along with the genera Hydrops and Pseudoeryx. The current work characterizes the proteolytic and neurotoxic activities of H. angulatus crude toxins from salivary excretion (SE) and describes the isolation and identification of a cysteine-rich secretory protein (CRISP) called helicopsin. The SE lethal dose (LD50) was 5.3 mg/kg; however, the SE did not contain hemorrhagic activity. Helicopsin was purified using activity-guided, Superose 12 10/300 GL molecular exclusion, Mono Q10 ion exchange, and Protein Pak 60 molecular exclusion. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) showed a highly purified band of approximately 20 kDa. The minimal lethal dose for helicopsin was 0.4 mg/kg. Liquid chromatography mass spectrometry (LC-MS/MS) analysis identified 2 unique peptides MEWYPEAAANAER and YTQIVWYK, representing a protein sequence (deleted homology) belonging to cysteine-rich secretory proteins, which are conserved in snake venoms (CRISPs). CRISPs are a large family of cysteine-rich secretory proteins found in various organisms and participate in diverse biological processes. Helicopsin exhibited robust neurotoxic activity as evidenced by immediate death (~8 min) due to respiratory paralysis in NIH mice. These observations for helicopsin purified from H. angulatus provide further evidence of the extensive distribution of highly potent neurotoxins in the Colubroidea superfamily of snakes than previously described. |
| Databáze: | OpenAIRE |
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