Enrichment of multiphosphorylated peptides by immobilized metal affinity chromatography using Ga(III)- and Fe(III)-complexes.
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.
Sykora
Corinna
C
Institute of Bioanalytical Chemistry, Centre for Biotechnology and Biomedicine (BBZ), Faculty of Chemistry and Mineralogy, Leipzig University, Deutscher Platz 5, 04103 Leipzig, Germany.
Hoffmann
Ralf
R
Hoffmann
Peter
P
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
D002364:Caseins; D005290:Ferric Compounds; D010748:Phosphopeptides; D011933:Reagent Kits, Diagnostic; D016875:tau Proteins; D010047:Ovalbumin; D005708:Gallium; D014357:Trypsin