title: Proteomic analysis of the rare Uracoan rattlesnake Crotalus vegrandis venom: Evidence of a broad arsenal of toxins.
abstract: The investigation of venoms has many clinical, pharmacological, ecological and evolutionary outcomes. The Crotalus spp. venom can cause hemorrhage, neurotoxicity, myotoxicity, coagulopathy and hypotension. Although neurotoxicity and hemorrhage usually does not occur for the same species, the rare Venezuelan species Crotalus vegrandis presents both characteristic. Different from the other species it has a restricted ecological niche and geographical distribution. Nevertheless, it has a raising medical importance as this rattlesnake population is increasing. Few works describe its neurotoxic and hemorrhagic features, but other toxins might play an important role in envenomation. We combined proteomic methods to identify for the first time the main components of it venom: 2D SDS-PAGE and gel-filtration chromatography for protein mixture decomplexation; LC-MS(2) of low molecular mass fractions and tryptic peptides; bioinformatic identification of toxin families and specific protein species based on unique peptide analysis and sequence database enriched with species-specific venom gland transcripts; and finally polyclonal anti-crotamine Western-blotting. Our results point to a broad arsenal of toxins in C. vegrandis venom: PIII and PII metalloproteases, crotoxin subunits, other phospholipases, isoforms of serine proteases and lectins, l-amino-acid oxidase, nerve growth factor, as well as other less abundant toxins.
authors.affiliation: Centro de Biotecnologia, IPEN, Av. Lineu Prestes 2242, CEP 05508-000, São Paulo, SP, Brazil.
authors.forename: Vincent Louis
authors.initials: VL
authors.lastname: Viala
authors.affiliation: Institut für Klinische Chemie, Universitätsklinikum Hamburg-Eppendorf, Haus Ost 26, Martinistr. 52, 20246, Hamburg, Germany.
authors.forename: Diana
authors.initials: D
authors.lastname: Hildebrand
authors.affiliation: Centro de Biotecnologia, IPEN, Av. Lineu Prestes 2242, CEP 05508-000, São Paulo, SP, Brazil.
authors.forename: Tamara Mieco
authors.initials: TM
authors.lastname: Fucase
authors.affiliation: Laboratório de Bioquímica e Biofísica, Instituto Butantan, Av. Vital Brasil, 1500, 05503-900, São Paulo, SP, Brazil.
authors.forename: Juliana Mozer
authors.initials: JM
authors.lastname: Sciani
authors.affiliation: Centro de Biotecnologia, IPEN, Av. Lineu Prestes 2242, CEP 05508-000, São Paulo, SP, Brazil.
authors.forename: José Pedro
authors.initials: JP
authors.lastname: Prezotto-Neto
authors.affiliation: Institut für Organische Chemie, Massenspektrometrie, Universität Hamburg, Martin-Luther-King-Platz 6, 20146, Hamburg, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Riedner
authors.affiliation: Laboratório Especial de Ecologia e Evolução, Instituto Butantan, Av. Dr. Vital Brasil 1500, CEP 05503-900, São Paulo, SP, Brazil.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Sanches
authors.affiliation: Centro de Biotecnologia, IPEN, Av. Lineu Prestes 2242, CEP 05508-000, São Paulo, SP, Brazil.
authors.forename: Paula Juliana
authors.initials: PJ
authors.lastname: Nishimura
authors.affiliation: Laboratório Especial de Ecologia e Evolução, Instituto Butantan, Av. Dr. Vital Brasil 1500, CEP 05503-900, São Paulo, SP, Brazil.
authors.forename: Nancy
authors.initials: N
authors.lastname: Oguiura
authors.affiliation: Laboratório de Bioquímica e Biofísica, Instituto Butantan, Av. Vital Brasil, 1500, 05503-900, São Paulo, SP, Brazil.
authors.forename: Daniel Carvalho
authors.initials: DC
authors.lastname: Pimenta
authors.affiliation: Institut für Klinische Chemie, Universitätsklinikum Hamburg-Eppendorf, Haus Ost 26, Martinistr. 52, 20246, Hamburg, Germany.
authors.forename: Hartmut
authors.initials: H
authors.lastname: Schlüter
authors.affiliation: Institut für Biochemie und Molekularbiologie, Universität Hamburg, Martin-Luther-King-Platz 6, 20146, Hamburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Betzel
authors.affiliation: Departamento de Física, IBILCE, UNESP, Rua Cristóvão Colombo, 2265, CEP 15054-000, São José do Rio Preto, SP, Brazil.
authors.forename: Raghuvir Krishnaswami
authors.initials: RK
authors.lastname: Arni
authors.affiliation: Centro de Biotecnologia, IPEN, Av. Lineu Prestes 2242, CEP 05508-000, São Paulo, SP, Brazil. Electronic address: pspencer@ipen.br.
authors.forename: Patrick Jack
authors.initials: PJ
authors.lastname: Spencer
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000737:chemistry*; D017839:Crotalus / Q000378:metabolism*; D013058:Mass Spectrometry; D045726:Metalloproteases / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D040901:Proteomics; D030162:Reptilian Proteins / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D003435:Crotalid Venoms; D030162:Reptilian Proteins; D045726:Metalloproteases
keywords: Crotalus vegrandis; Peptide; Proteomics; Toxin; Uracoan rattlesnake; Venom