title: A cleavable biotin tagging reagent that enables the enrichment and identification of carbonylation sites in proteins.
abstract: The utility of a new, cleavable tag for identifying and enriching protein carbonyls is examined. Using a model system, human serum albumin modified with acrolein, the EZ-Link alkoxyamine-PEG4-SS-PEG4-biotin affinity tag, was tested for its ability to label protein carbonyls in proteomic analyses of protein carbonylation. The efficiency of the labeling was assayed and compared to standard biotin hydrazide reagents. The label was also tested in liquid chromatography-tandem mass spectrometry (LC/MS/MS) experiments. The quality of the fragmentation spectra was assessed and the relative detection efficiency of various modification sites was compared to standard biotin hydrazide reagents. Finally, the viability of using the label with streptavidin bead enrichment protocols in a standard proteomics workflow was probed.
authors.affiliation: Department of Chemistry, Virginia Commonwealth University, 1001 West Main Street, P.O. Box 842006, Richmond, VA, 23284-2006, USA.
authors.forename: Chelsea M
authors.initials: CM
authors.lastname: Coffey
authors.affiliation: Department of Chemistry, Virginia Commonwealth University, 1001 West Main Street, P.O. Box 842006, Richmond, VA, 23284-2006, USA. sgronert@vcu.edu.
authors.forename: Scott
authors.initials: S
authors.lastname: Gronert
mesh_terms: D001710:Biotin / Q000031:analogs & derivatives / Q000737:chemistry; D006801:Humans; D050050:Protein Carbonylation; D040901:Proteomics / Q000379:methods*; D012709:Serum Albumin / Q000737:chemistry*
chemical_list: D012709:Serum Albumin; C053311:biotin hydrazide; D001710:Biotin
keywords: Acrolein; Mass spectrometry; Oxidative stress; Protein carbonylation; Proteomics