title: Enrichment of multiphosphorylated peptides by immobilized metal affinity chromatography using Ga(III)- and Fe(III)-complexes.
abstract: The detection and identification of O-phosphorylation sites in proteins with mass spectrometry remains a challenge. A common approach to analyse these modifications is to enrich phosphopeptides by immobilized metal affinity chromatography (IMAC) prior to mass spectrometric analysis. In this study two commercially available IMAC kits based on Fe(III)-ions immobilized on magnetic beads and Ga(III)-ions immobilized on a chelate-resin, have been investigated and the binding efficiency of peptide mixtures containing non-phosphorylated, singly, doubly and triply phosphorylated peptides have been tested.
authors.affiliation: Institute of Bioanalytical Chemistry, Centre for Biotechnology and Biomedicine (BBZ), Faculty of Chemistry and Mineralogy, Leipzig University, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Corinna
authors.initials: C
authors.lastname: Sykora
authors.forename: Ralf
authors.initials: R
authors.lastname: Hoffmann
authors.forename: Peter
authors.initials: P
authors.lastname: Hoffmann
mesh_terms: D002364:Caseins / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods*; D005290:Ferric Compounds / Q000737:chemistry*; D005708:Gallium / Q000737:chemistry*; D006801:Humans; D010047:Ovalbumin / Q000302:isolation & purification; D010748:Phosphopeptides / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism; D016875:tau Proteins / Q000302:isolation & purification
chemical_list: D002364:Caseins; D005290:Ferric Compounds; D010748:Phosphopeptides; D011933:Reagent Kits, Diagnostic; D016875:tau Proteins; D010047:Ovalbumin; D005708:Gallium; D014357:Trypsin