title: Molecular profiling of the human nasal epithelium: A proteomics approach.
abstract: A comprehensive proteomic profiling of nasal epithelium (NE) is described. This study relies on simple subcellular fractionation used to obtain soluble- and membrane-enriched fractions followed by 2-dimensional liquid chromatography (2D-LC) separation and tandem mass spectrometry (MS/MS). The cells were collected using a brushing technique applied on NE of clinically evaluated volunteers. Subsequently, the soluble- and the membrane-protein enriched fractions were prepared and analyzed in parallel using 2D-LC-MS/MS. In a set of 1482 identified proteins, 947 (63.9%) proteins were found to be associated to membrane fraction. Grand average hydropathy value index (GRAVY) analysis, the transmembrane protein mapping and annotations of primary location deposited in the Human Protein Reference Database (HPRD) confirmed an enrichment of hydrophobic proteins on this dataset. Ingenuity Pathway Analysis (IPA) of soluble fraction revealed an enrichment of molecular and cellular functions associated with cell death, protein folding and drug metabolism while in membrane fraction showed an enrichment of functions associated with molecular transport, protein trafficking and cell-to-cell signaling and interaction. The IPA showed similar enrichment of functions associated with cellular growth and proliferation in both soluble and membrane subproteomes. This finding was in agreement with protein content analysis using exponentially modified protein abundance index (emPAI). A comparison of our data with previously published studies focusing on respiratory tract epithelium revealed similarities related to identification of proteins associated with physical barrier function and immunological defence. In summary, we extended the NE molecular profile by identifying and characterizing proteins associated to pivotal functions of a respiratory epithelium, including the control of fluid volume and ionic composition at the airways' surface, physical barrier maintenance, detoxification and immunological defence. The extent of similarities supports the applicability of a less invasive analysis of NE to assess prognosis and treatment response of lung diseases such as asthma, cystic fibrosis and chronic obstructive pulmonary disease.
authors.affiliation: Laboratório de Proteómica, Departamento de Genética, Instituto Nacional de Saúde Dr. Ricardo Jorge, INSA I.P., Portugal.
authors.forename: Tânia
authors.initials: T
authors.lastname: Simões
authors.forename: Nuno
authors.initials: N
authors.lastname: Charro
authors.forename: Josip
authors.initials: J
authors.lastname: Blonder
authors.forename: Daniel
authors.initials: D
authors.lastname: Faria
authors.forename: Francisco M
authors.initials: FM
authors.lastname: Couto
authors.forename: King C
authors.initials: KC
authors.lastname: Chan
authors.forename: Timothy
authors.initials: T
authors.lastname: Waybright
authors.forename: Haleem J
authors.initials: HJ
authors.lastname: Isaaq
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Veenstra
authors.forename: Deborah
authors.initials: D
authors.lastname: Penque
mesh_terms: D002458:Cell Fractionation / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry; D009297:Nasal Mucosa / Q000737:chemistry* / Q000166:cytology; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D008565:Membrane Proteins