title: Identification of selenium-containing proteins in HEK 293 kidney cells using multiple chromatographies, LC-ICPMS and nano-LC-ESIMS.
abstract: Our previous studies using HeLa and HEK 293 cells demonstrated that selenomethionine, SeMet, exerts more of an antagonistic effect on arsenic than other selenium species. These studies attributed the antagonistic effect of SeMet to decreased levels of reactive oxygen species, ROS, changes in protein phosphorylation and possible incorporation of SeMet into proteins. The present study employs a metallomics approach to identify the selenium-containing proteins in HEK 293 cells raised with SeMet. The proteins were screened and separated using two dimensional high performance liquid chromatography (HPLC)-inductively coupled plasma mass spectrometry (ICPMS), size exclusion chromatography (SEC) and reversed-phase chromatography (RPC). The Se-containing proteins were identified by peptide mapping using nano-HPLC-Chip-electrospray ionization mass spectrometry (ESIMS).
authors: University of Cincinnati/Agilent Technologies Metallomics Center of the Americas, Department of Chemistry, University of Cincinnati, Cincinnati, OH 45221-0172, USA.
authors: Karnakar R
authors: KR
authors: Chitta
authors: Julio A
authors: JA
authors: Landero-Figueroa
authors: Phanichand
authors: P
authors: Kodali
authors: Joseph A
authors: JA
authors: Caruso
authors: Edward J
authors: EJ
authors: Merino
mesh_terms: D002845:Chromatography / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011506:Proteins / Q000032:analysis / Q000737:chemistry*; D012643:Selenium / Q000032:analysis*; D012645:Selenomethionine / Q000494:pharmacology; D020539:Sequence Analysis, Protein
chemical_list: D011506:Proteins; D012645:Selenomethionine; D012643:Selenium
keywords: Identifying selenium-containing proteins; LC–ESIMS; Metallomics; Multiple chromatographic separations; SEC–ICPMS