title: Application of iTRAQ Shotgun Proteomics for Measurement of Brain Proteins in Studies of Psychiatric Disorders.
abstract: Shotgun proteomics has been used successfully for more than two decades for measurement of proteins from diverse biological systems. This has led to new insights and identification of potential biomarkers and drug targets for numerous medical conditions. The advent of mass-labeling approaches such as isobaric tagging for relative and absolute quantitation (iTRAQ) has allowed multiplexing of samples to improve the accuracy and throughput of experiments and the reduction of costs. Here, we describe a detailed iTRAQ mass spectrometry protocol for simultaneous analysis of four proteomes from postmortem brain tissue extracts. In addition, a post-labeling procedure for peptide fractionation is presented.
authors.affiliation: Laboratory of Protein Chemistry - Proteomics Unit, Chemistry Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Erika Velásquez
authors.initials: EV
authors.lastname: Núñez
authors.affiliation: Department of Biochemistry and Tissue Biology, Laboratory of Neuroproteomics, Institute of Biology, University of Campinas, CidadeUniversitáriaZeferinoVaz, Campinas, Brazil.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Guest
authors.affiliation: Department of Biochemistry and Tissue Biology, Laboratory of Neuroproteomics, Institute of Biology, University of Campinas, CidadeUniversitáriaZeferinoVaz, Campinas, Brazil.
authors.forename: Daniel
authors.initials: D
authors.lastname: Martins-de-Souza
authors.affiliation: Laboratory of Protein Chemistry - Proteomics Unit, Chemistry Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Gilberto Barbosa
authors.initials: GB
authors.lastname: Domont
authors.affiliation: Laboratory of Protein Chemistry - Proteomics Unit, Chemistry Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil. fabiocsn@ufrj.br.
authors.forename: Fábio César Sousa
authors.initials: FC
authors.lastname: Nogueira
mesh_terms: D015415:Biomarkers / Q000032:analysis; D001923:Brain Chemistry; D002853:Chromatography, Liquid / Q000379:methods; D006801:Humans; D001523:Mental Disorders / Q000378:metabolism*; D008970:Molecular Weight; D036103:Nanotechnology / Q000379:methods; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000302:isolation & purification; D010446:Peptide Fragments / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013194:Staining and Labeling / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D015415:Biomarkers; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments
keywords: Brain tissues; Mass spectrometry; Mass tagging; Proteomics; Psychiatric disorders; iTRAQ