caa a22 4500 445853719 CHVBK 20180317145421.0 cr unu---uuuuu 170323e20110401xx s 000 0 eng 10.1007/s00204-010-0597-6 doi (NATIONALLICENCE)springer-10.1007/s00204-010-0597-6 Characterization of toxins from the broad-banded water snake Helicops angulatus (Linnaeus, 1758): isolation of a cysteine-rich secretory protein, Helicopsin [Elektronische Daten] [Amalid Estrella, Elda Sánchez, Jacob Galán, W. Tao, Belsy Guerrero, Luis Navarrete, Alexis Rodríguez-Acosta] 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 5.3mg/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 20kDa. The minimal lethal dose for helicopsin was 0.4mg/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 (~8min) 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. Springer-Verlag, 2010 CRISP nationallicence Colubroidea nationallicence Dipsadidae salivary excretion nationallicence Neurotoxin nationallicence Helicops angulatus nationallicence Broad-banded water snake nationallicence Estrella Amalid Immunochemistry Section, Tropical Medicine Institute of the Universidad Central de Venezuela, Apdo. 47423, 1041, Caracas, Venezuela aut Sánchez Elda Department of Chemistry, College of Arts and Sciences, Texas A&M University-Kingsville, MSC 158, 78363, Kingsville, TX, USA aut Galán Jacob Departments of Biochemistry, Chemistry, Medicinal Chemistry & Molecular Pharmacology, Purdue University, 47907, West Lafayette, IN, USA aut Tao W. Departments of Biochemistry, Chemistry, Medicinal Chemistry & Molecular Pharmacology, Purdue University, 47907, West Lafayette, IN, USA aut Guerrero Belsy Pathophysiology Laboratory, Medicine Experimental Center, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, Venezuela aut Navarrete Luis Immunochemistry Section, Tropical Medicine Institute of the Universidad Central de Venezuela, Apdo. 47423, 1041, Caracas, Venezuela aut Rodríguez-Acosta Alexis Immunochemistry Section, Tropical Medicine Institute of the Universidad Central de Venezuela, Apdo. 47423, 1041, Caracas, Venezuela aut Archives of Toxicology Springer-Verlag 85/4(2011-04-01), 305-313 0340-5761 85:4<305 2011 85 204 https://doi.org/10.1007/s00204-010-0597-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00204-010-0597-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Estrella Amalid Immunochemistry Section, Tropical Medicine Institute of the Universidad Central de Venezuela, Apdo. 47423, 1041, Caracas, Venezuela aut NATIONALLICENCE 700 1- Sánchez Elda Department of Chemistry, College of Arts and Sciences, Texas A&M University-Kingsville, MSC 158, 78363, Kingsville, TX, USA aut NATIONALLICENCE 700 1- Galán Jacob Departments of Biochemistry, Chemistry, Medicinal Chemistry & Molecular Pharmacology, Purdue University, 47907, West Lafayette, IN, USA aut NATIONALLICENCE 700 1- Tao W. Departments of Biochemistry, Chemistry, Medicinal Chemistry & Molecular Pharmacology, Purdue University, 47907, West Lafayette, IN, USA aut NATIONALLICENCE 700 1- Guerrero Belsy Pathophysiology Laboratory, Medicine Experimental Center, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, Venezuela aut NATIONALLICENCE 700 1- Navarrete Luis Immunochemistry Section, Tropical Medicine Institute of the Universidad Central de Venezuela, Apdo. 47423, 1041, Caracas, Venezuela aut NATIONALLICENCE 700 1- Rodríguez-Acosta Alexis Immunochemistry Section, Tropical Medicine Institute of the Universidad Central de Venezuela, Apdo. 47423, 1041, Caracas, Venezuela aut NATIONALLICENCE 773 0- Archives of Toxicology Springer-Verlag 85/4(2011-04-01), 305-313 0340-5761 85:4<305 2011 85 204 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer