title: Pseudechis guttatus venom proteome: Insights into evolution and toxin clustering.
abstract: The Australian Elapidae spotted black snake Pseudechis guttatus venom proteome composition was analyzed by high throughput mass spectrometry. The crude venom proteins were decomplexed by 2D-PAGE and in-gel digestion peptides from 66 spot samples and analyzed by tandem mass spectrometry-LC-ESI-ion trap. Protein identification was performed combining PEAKS studio 7.0 and Mascot software. The analysis identified l-amino-acid oxidases, phospholipases A2, metalloproteases, nerve growth factors and ecto-5'-nucleotidases, and for the first time in this venom the components cysteine-rich secretory proteins similar to pseudechetoxin, phospholipase B and transferrin-like protein. The envenomation symptoms are in agreement with the identified components, but the present limitations of database information might impair the detection of toxin families, protein species and still unknown toxins. From the qualitative point of view, the similarity of this venom with the ones from other Pseudechis species could be assigned to recent speciation events.
  
  
  CONCLUSIONS
  Studies on the proteome of Australian Elapidae (Ancanthophiinae) are quite rare. In the present work we performed, using classic proteomic methods, a qualitative and partial analysis of the proteic components of Pseudechis guttatus venom. Although previous studies contributed to the knowledge of the major components of this venom, our study revealed some yet undescribed protein species, as well as new toxins, such as CRiSPs, phospholipase B, transferrin-like protein and ecto 5'-nucleotidase.
authors.affiliation: Centro de Biotecnologia, IPEN, Av. Lineu Prestes 2242, 05508-000 São Paulo, SP, Brazil. Electronic address: pspencer@ipen.br.
authors.forename: Vincent L
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: Institut für Klinische Chemie, Universitätsklinikum Hamburg-Eppendorf, Haus Ost 26, Martinistr. 52, 20246 Hamburg, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Trusch
authors.affiliation: Departamento de Física, IBILCE, UNESP, Rua Cristóvão Colombo, 2265, 15054-000 São José do Rio Preto, SP, Brazil.
authors.forename: Raghuvir K
authors.initials: RK
authors.lastname: Arni
authors.affiliation: Laboratório de Bioquímica e Biofísica, Instituto Butantan, Av. Vital Brasil, 1500, 05503-900 São Paulo, SP, Brazil.
authors.forename: Daniel C
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: Centro de Biotecnologia, IPEN, Av. Lineu Prestes 2242, 05508-000 São Paulo, SP, Brazil.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Spencer
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D005075:Biological Evolution; D004546:Elapid Venoms / Q000737:chemistry* / Q000235:genetics*; D017815:Elapidae / Q000145:classification / Q000235:genetics*; D019143:Evolution, Molecular; D014644:Genetic Variation; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics*; D020543:Proteome / Q000235:genetics*; D020539:Sequence Analysis, Protein / Q000379:methods; D013045:Species Specificity
chemical_list: D004546:Elapid Venoms; D020543:Proteome; C026235:Pseudechis venom
keywords: 2D-PAGE; Elapidae; High throughput; Proteomics; Pseudechis guttatus; Toxins