title: Selenium effects on arsenic cytotoxicity and protein phosphorylation in human kidney cells using chip-based nanoLC-MS/MS.
abstract: Although arsenic toxicity is well known, little is known of how it exerts its effects at the proteome level. Protein phosphorylation is an important post-translational modification in the regulation of cell signaling. Despite the importance of protein phosphorylation, the identification and characterization of phosphorylated proteins, as influenced by interaction between arsenic and selenium species have not been fully studied. The aim of this study is to identify phosphorylation in arsenic toxified cells, with and without selenium present. Here, we identify the phosphorylated proteins related to post translational modifications (PTMs) after inorganic arsenic (iAs) and selenomethionine (SeMet) were inoculated together with HEK293 human kidney cells. In this study, using TiO(2)-based nanoLC-phosphochip® coupled to ESI-MS we observed phosphorylated peptide enrichment and significant reduction in sample complexity. The identification of phosphorylated proteins in highly complex digests of cell lysate were markedly different with As toxification only, or when in the presence of SeMet. Several phosphorylation sites and proteins are identified using Spectrum Mill and Mascot protein data base search engines. Cytotoxicity studies showed that SeMet significantly reduces the cytotoxic effect of iAs in HEK293 cells, while inorganic selenium did not.
authors.affiliation: Analytical Chemistry Department, Faculty of Pharmacy, Gazi University, Ankara, Turkey.
authors.forename: Orkun
authors.initials: O
authors.lastname: Alp
authors.forename: Yaofang
authors.initials: Y
authors.lastname: Zhang
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Merino
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Caruso
mesh_terms: D000595:Amino Acid Sequence; D001151:Arsenic / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D002853:Chromatography, Liquid; D057809:HEK293 Cells; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D008969:Molecular Sequence Data; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods*; D010750:Phosphoproteins / Q000032:analysis / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011506:Proteins / Q000378:metabolism*; D020543:Proteome / Q000032:analysis / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D012643:Selenium / Q000494:pharmacology*; D012645:Selenomethionine / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D010750:Phosphoproteins; D011506:Proteins; D020543:Proteome; D012645:Selenomethionine; D012643:Selenium; D001151:Arsenic