title: Proteomic analysis of Toxocara canis excretory and secretory (TES) proteins.
abstract: Toxocariasis is a neglected disease, and its main etiological agent is the nematode Toxocara canis. Serological diagnosis is performed by an enzyme-linked immunosorbent assay using T. canis excretory and secretory (TES) antigens produced by in vitro cultivation of larvae. Identification of TES proteins can be useful for the development of new diagnostic strategies since few TES components have been described so far. Herein, we report the results obtained by proteomic analysis of TES proteins using a liquid chromatography-tandem mass spectrometry (LC-MS/MS) approach. TES fractions were separated by one-dimensional SDS-PAGE and analyzed by LC-MS/MS. The MS/MS spectra were compared with a database of protein sequences deduced from the genome sequence of T. canis, and a total of 19 proteins were identified. Classification according to the signal peptide prediction using the SignalP server showed that seven of the identified proteins were extracellular, 10 had cytoplasmic or nuclear localization, while the subcellular localization of two proteins was unknown. Analysis of molecular functions by BLAST2GO showed that the majority of the gene ontology (GO) terms associated with the proteins present in the TES sample were associated with binding functions, including but not limited to protein binding (GO:0005515), inorganic ion binding (GO:0043167), and organic cyclic compound binding (GO:0097159). This study provides additional information about the exoproteome of T. canis, which can lead to the development of new strategies for diagnostics or vaccination.
authors.affiliation: Departamento de Microbiologia e Parasitologia, Universidade Federal de Pelotas (UFPel), Pelotas, Brazil.
authors.forename: Rita Leal
authors.initials: RL
authors.lastname: Sperotto
authors.affiliation: Laboratório de Biotecnologia Infecto-Parasitária, Centro de Desenvolvimento Tecnológico- UFPel, Pelotas, Brazil.
authors.forename: Frederico Schmitt
authors.initials: FS
authors.lastname: Kremer
authors.affiliation: Departamento de Microbiologia e Parasitologia, Universidade Federal de Pelotas (UFPel), Pelotas, Brazil.
authors.forename: Maria Elisabeth
authors.initials: ME
authors.lastname: Aires Berne
authors.affiliation: Departamento de Microbiologia e Parasitologia, Universidade Federal de Pelotas (UFPel), Pelotas, Brazil.
authors.forename: Luciana F
authors.initials: LF
authors.lastname: Costa de Avila
authors.affiliation: Laboratório de Biotecnologia Infecto-Parasitária, Centro de Desenvolvimento Tecnológico- UFPel, Pelotas, Brazil.
authors.forename: Luciano
authors.initials: L
authors.lastname: da Silva Pinto
authors.affiliation: Laboratório de Genômica Estrutural e Funcional, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil; Departamento de Biologia Molecular e Biotecnologia, Instituto de Biociências, UFRGS, Porto Alegre, Brazil.
authors.forename: Karina Mariante
authors.initials: KM
authors.lastname: Monteiro
authors.affiliation: Laboratório de Genômica Estrutural e Funcional, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil; Departamento de Biologia Molecular e Biotecnologia, Instituto de Biociências, UFRGS, Porto Alegre, Brazil.
authors.forename: Karin Silva
authors.initials: KS
authors.lastname: Caumo
authors.affiliation: Laboratório de Genômica Estrutural e Funcional, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil; Departamento de Biologia Molecular e Biotecnologia, Instituto de Biociências, UFRGS, Porto Alegre, Brazil.
authors.forename: Henrique Bunselmeyer
authors.initials: HB
authors.lastname: Ferreira
authors.affiliation: Departamento de Microbiologia e Parasitologia, Universidade Federal de Pelotas (UFPel), Pelotas, Brazil.
authors.forename: Natália
authors.initials: N
authors.lastname: Berne
authors.affiliation: Laboratório de Biotecnologia Infecto-Parasitária, Centro de Desenvolvimento Tecnológico- UFPel, Pelotas, Brazil. Electronic address: sibeleborsuk@gmail.com.
authors.forename: Sibele
authors.initials: S
authors.lastname: Borsuk
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D019295:Computational Biology / Q000379:methods; D004285:Dogs; D055354:Exosomes / Q000378:metabolism*; D005260:Female; D015801:Helminth Proteins / Q000378:metabolism*; D020543:Proteome; D040901:Proteomics / Q000379:methods; D022142:Secretory Vesicles / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D017166:Toxocara canis / Q000378:metabolism*; D014120:Toxocariasis / Q000469:parasitology
chemical_list: D015801:Helminth Proteins; D020543:Proteome
keywords: LC–MS/MS; Proteomic analysis; TES; Toxocara canis