title: Profiling Cell Lines Nuclear Sub-proteome.
abstract: Proteins are very dynamic within the cell and their localization and trafficking between subcellular compartments are critical for their correct function. Indeed, the abnormal localization of a protein might lead to the pathogenesis of several diseases. The association of cell fractionation methods and mass spectrometry based proteomic methods allow both the localization and quantification of proteins in different sub-compartments. Here we present a detailed protocol for enrichment, identification, and quantitation of the nuclear proteome in cell lines combining nuclear subproteome enrichment by differential centrifugation and high-throughput proteomics.
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil.|Center for Cell Based Therapy - Hemotherapy Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Aline
authors.initials: A
authors.lastname: Poersch
authors.affiliation: Department of Pediatrics, Ribeirão Preto Medical School, University of São Paulo, Rua Tenente Catão Roxo, 2.501, Ribeirão Preto, 14051-140, SP, Brazil.
authors.forename: Andrea G
authors.initials: AG
authors.lastname: Maria
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil.|Center for Cell Based Therapy - Hemotherapy Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Camila S
authors.initials: CS
authors.lastname: Palma
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil.|Center for Cell Based Therapy - Hemotherapy Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Mariana L
authors.initials: ML
authors.lastname: Grassi
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Daniele
authors.initials: D
authors.lastname: Albuquerque
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil.|Center for Cell Based Therapy - Hemotherapy Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Carolina H
authors.initials: CH
authors.lastname: Thomé
authors.affiliation: Department of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil. vitor.faca@fmrp.usp.br.|Center for Cell Based Therapy - Hemotherapy Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil. vitor.faca@fmrp.usp.br.
authors.forename: Vitor M
authors.initials: VM
authors.lastname: Faça
mesh_terms: D002458:Cell Fractionation / Q000379:methods; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D019295:Computational Biology / Q000379:methods; D030562:Databases, Protein; D057166:High-Throughput Screening Assays; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D009687:Nuclear Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D021381:Protein Transport; D020543:Proteome; D040901:Proteomics / Q000379:methods*
chemical_list: D009687:Nuclear Proteins; D020543:Proteome
keywords: Cell Line; Mass spectrometry; Nuclear fractionation; Protein localization; Subcellular proteomics