title: HCD Fragmentation of Glycated Peptides.
abstract: Protein glycation is a concentration-dependent nonenzymatic reaction of reducing sugars with amine groups of proteins to form early as well as advanced glycation (end-) products (AGEs). Glycation is a highly disease-relevant modification but is typically only studied on a few blood proteins. To complement our blood proteomics studies in diabetics, we here investigate protein glycation by higher energy collisional dissociation (HCD) fragmentation on Orbitrap mass spectrometers. We established parameters to most efficiently fragment and identify early glycation products on in vitro glycated model proteins. Retaining standard collision energies does not degrade performance if the most dominant neutral loss of H6O3 is included into the database search strategy. Glycation analysis of the entire HeLa proteome revealed an unexpected intracellular preponderance for arginine over lysine modification in early and advanced glycation (end-) products. Single-run analysis from 1 μL of undepleted and unenriched blood plasma identified 101 early glycation sites as well as numerous AGE sites on diverse plasma proteins. We conclude that HCD fragmentation is well-suited for analyzing glycated peptides and that the diabetic status of patients can be directly diagnosed from single-run plasma proteomics measurements.
authors.affiliation: Department of Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry , Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Eva C
authors.initials: EC
authors.lastname: Keilhauer
authors.affiliation: Department of Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry , Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Philipp E
authors.initials: PE
authors.lastname: Geyer
authors.affiliation: Department of Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry , Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Mann
mesh_terms: D001120:Arginine / Q000737:chemistry; D003920:Diabetes Mellitus / Q000175:diagnosis; D017127:Glycation End Products, Advanced / Q000378:metabolism; D006020:Glycopeptides / Q000032:analysis* / Q000737:chemistry; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D008239:Lysine / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D010446:Peptide Fragments / Q000737:chemistry*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics
chemical_list: D017127:Glycation End Products, Advanced; D006020:Glycopeptides; D010446:Peptide Fragments; D020543:Proteome; D001120:Arginine; D008239:Lysine
keywords: AGEs; blood plasma; diabetes; higher-energy collisional dissociation; protein glycation