title: Characterization of selenium incorporation into wheat proteins by two-dimensional gel electrophoresis-laser ablation ICP MS followed by capillary HPLC-ICP MS and electrospray linear trap quadrupole Orbitrap MS.
abstract: A method has been developed for a rapid and precise location of selenium-containing proteins in large two-dimensional (2D) electrophoresis gels. A sample was divided into four aliquots which were analyzed in parallel by 1D isoelectric focusing electrophoresis (IEF)-laser ablation (LA) inductively coupled plasma mass spectrometry (ICP MS), 1D sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE)-LA ICP MS, and, in duplicate, by 2D IEF-PAGE. On the basis of the 1 D electropherograms obtained, areas supposed to contain the largest concentrations of Se were subjected to LA ICP MS imaging to locate precisely the position of Se-containing proteins which were then identified in the parallel 2D gel by electrospray Orbitrap MS/MS. The method was applied to the identification and semiquantitative determination of selenium storage proteins in wheat. MS evidence is presented for the Se-S substitution in plants not only in methionine but also in cysteine.
authors: CNRS-UPPA, Laboratoire de Chimie Analytique Bio-inorganique et Environnement, UMR5254, Hélioparc, 2, Av. Angot, 64053 Pau, France.
authors: Juliusz
authors: J
authors: Bianga
authors: Espen
authors: E
authors: Govasmark
authors: Joanna
authors: J
authors: Szpunar
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation; D003545:Cysteine / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D007525:Isoelectric Focusing; D008715:Methionine / Q000378:metabolism; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D010940:Plant Proteins / Q000032:analysis* / Q000378:metabolism; D012643:Selenium / Q000032:analysis* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation / Q000379:methods*; D014908:Triticum / Q000378:metabolism*
chemical_list: D010455:Peptides; D010940:Plant Proteins; D008715:Methionine; D012643:Selenium; D003545:Cysteine