Venom characterization of the three species of Ophryacus and proteomic profiling of O-sphenophrys unveils Sphenotoxin, a novel Crotoxin-like heterodimeric beta-neurotoxin
Por:
Neri-Castro E., Lomonte B., Valdés M., Ponce-López R., Bénard-Valle M., Borja M., Strickland J.L., Jones J.M., Grünwald C., Zamudio F., Alagón A.
Publicada:
10 feb 2019
Ahead of Print:
1 ene 2018
Resumen:
Venoms of the three species of Ophryacus (O. sphenophrys, O.
smaragdinus, and O. undulatus), a viperid genus endemic to Mexico, were
analyzed for the first time in the present work. The three venoms lacked
procoagulant activity on human plasma, but induced hemorrhage and were
highly lethal to mice. These venoms also displayed proteolytic and
phospholipase A(2) activities in vitro. The venom of O. sphenophrys was
the most lethal and caused hind-limb paralysis in mice. Proteomic
profiling of O. sphenophrys venom showed a predominance of
metalloproteinase (34.9%), phospholipase A(2) (24.8%) and serine
protease (17.1%) in its composition. Strikingly, within its PLA(2)
components, 12.9% corresponded to a Crotoxin-like heterodimer, here
named Sphenotoxin, which was not found in the other two species of
Ophryacus. Sphenotoxin, like Crotoxin, is composed of non-covalently
bound A and B subunits. Partial amino acid sequence was obtained for
Sphenotoxin B and was similar (78-89%) to other subunits described. The
mouse i.v. LD50 of Sphenotoxin at 1:1 M radio was 0.16 mu g/g. Also,
like Crotoxin, Sphenotoxin induced a potent neuromuscular blockade in
the phrenic nerve-diaphragm preparation. Ophryacus is the fifth genus
and 0. sphenophrys the third non-rattlesnake species shown to contain a
novel Crotoxin-like heterodimeric beta-neurotoxin.
Biological significance: Ophryacus is an endemic genus of semi-arboreal
pitvipers from Mexico that includes three species with restricted
distributions. Little is known about the natural history of these
species and nothing is known about the properties of their venoms.
Research on these species' venoms could generate relevant information
regarding venom composition of Mexican pitvipers. Additionally, research
into the presence of neurotoxic Crotoxin-like molecules outside of
rattlesnakes (genera Crotalus and Sistrurus) has identified this
molecule in several new genera. Knowing which genera and species possess
neurotoxic components is important to fully understand the repercussions
of snakebites, the interaction with prey and predators, and the origin,
evolution, and phylogenetic distribution of Crotoxin-like molecules
during the evolutionary history of pitvipers.
Our study expands current knowledge regarding venom's compositions and
function from Mexican pitvipers, providing a comparative venom
characterization of major activities in the three Ophryacus species.
Additionally, the discovery and characterization of a novel
Crotoxin-like molecule, here named Sphenotoxin, in O. sphenophrys, and
the detailed protein composition of O. sphenophrys venom supports the
hypotheses that Crotoxin-like-beta-neurotoxins are more widespread than
initially thought.
Filiaciones:
Neri-Castro E.:
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Colonia Chamilpa, Cuernavaca, Morelos 62210, Mexico
Programa de Doctorado en Ciencias Biomédicas UNAM, Mexico
Univ Nacl Autonoma Mexico, Inst Biotecnol, Av Univ 2001, Cuernavaca 62210, Morelos, Mexico
Univ Nacl Autonoma Mexico, Programa Doctorado Ciencias Biomed, Mexico City, DF, Mexico
Lomonte B.:
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, 11501, Costa Rica
Univ Costa Rica, Fac Microbiol, Inst Clodomiro Picado, San Jose 11501, Costa Rica
Valdés M.:
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Colonia Chamilpa, Cuernavaca, Morelos 62210, Mexico
Laboratorio de Genética, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional07738, Mexico
Univ Nacl Autonoma Mexico, Inst Biotecnol, Av Univ 2001, Cuernavaca 62210, Morelos, Mexico
Inst Politecn Nacl, Escuela Nacl Ciencias Biol, Lab Genet, Mexico City 07738, DF, Mexico
Ponce-López R.:
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Colonia Chamilpa, Cuernavaca, Morelos 62210, Mexico
Univ Nacl Autonoma Mexico, Inst Biotecnol, Av Univ 2001, Cuernavaca 62210, Morelos, Mexico
Bénard-Valle M.:
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Colonia Chamilpa, Cuernavaca, Morelos 62210, Mexico
Univ Nacl Autonoma Mexico, Inst Biotecnol, Av Univ 2001, Cuernavaca 62210, Morelos, Mexico
Borja M.:
Facultad de Ciencias Biológicas, Universidad Juárez del Estado de Durango, Av. Universidad s/n. Fracc. Filadelfia, Gómez Palacio, 35010, Mexico
Univ Juarez Estado Durango, Fac Ciencias Biol, Av Univ S-N, Fracc Filadelfia 35010, Gomez Palacio, Mexico
Strickland J.L.:
Department of Biology, University of Central Florida, 4000 Central Florida Blvd, Orlando, FL 32816, United States
Univ Cent Florida, Dept Biol, 4000 Cent Florida Blvd, Orlando, FL 32816 USA
Jones J.M.:
Herp.mx A.C., Villa del Álvarez, Colima, Mexico
Herp Mx AC, Villa Del Alvarez, Colima, Mexico
Grünwald C.:
Herp.mx A.C., Villa del Álvarez, Colima, Mexico
Herp Mx AC, Villa Del Alvarez, Colima, Mexico
Zamudio F.:
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Colonia Chamilpa, Cuernavaca, Morelos 62210, Mexico
Univ Nacl Autonoma Mexico, Inst Biotecnol, Av Univ 2001, Cuernavaca 62210, Morelos, Mexico
Alagón A.:
Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Colonia Chamilpa, Cuernavaca, Morelos 62210, Mexico
Univ Nacl Autonoma Mexico, Inst Biotecnol, Av Univ 2001, Cuernavaca 62210, Morelos, Mexico
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