title: Fibrinogen alpha chain O-glycopeptides as possible markers of urinary tract infection.
abstract: Urinary tract infection (UTI) is the most common bacterial infection leading to substantial morbidity and considerable health care expenditures across all ages. Here we present an exploratory UPLC-MS study of human urine in the context of febrile, complicated urinary tract infection aimed to reveal and identify possible markers of a host response on infection. A UPLC-MS based workflow, taking advantage of Ultra High Resolution (UHR) Qq-ToF-MS, and multivariate data handling were applied to a carefully selected group of 39 subjects with culture-confirmed febrile Escherichia coli UTI. Using a combination of unsupervised and supervised multivariate modeling we have pinpointed a number of peptides specific for UTI. An unequivocal structural identification of these peptides, as O-glycosylated fragments of the human fibrinogen alpha 1 chain, required MS2 and MS3 experiments on two different MS platforms: ESI-UHR-Qq-ToF and ESI-ion trap, a blast search and, finally, confirmation was achieved by matching experimental tandem mass spectra with those of custom synthesized candidate-peptides. In conclusion, exploiting non-targeted UPLC-MS based approach for the investigation of UTI related changes in urine, we have identified and structurally characterized unique O-glycopeptides, which are, to our knowledge, the first demonstration of O-glycosylation of human fibrinogen alpha 1-chain.
authors.affiliation: Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands. T.Pacchiarotta@lumc.nl
authors.forename: Tiziana
authors.initials: T
authors.lastname: Pacchiarotta
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Hensbergen
authors.forename: Manfred
authors.initials: M
authors.lastname: Wuhrer
authors.forename: Cees
authors.initials: C
authors.lastname: van Nieuwkoop
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Nevedomskaya
authors.forename: Rico J E
authors.initials: RJ
authors.lastname: Derks
authors.forename: Bart
authors.initials: B
authors.lastname: Schoenmaker
authors.forename: Carolien A M
authors.initials: CA
authors.lastname: Koeleman
authors.forename: Jaap
authors.initials: J
authors.lastname: van Dissel
authors.forename: André M
authors.initials: AM
authors.lastname: Deelder
authors.forename: Oleg A
authors.initials: OA
authors.lastname: Mayboroda
mesh_terms: D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000652:urine; D004926:Escherichia coli; D004927:Escherichia coli Infections / Q000652:urine*; D005260:Female; D005340:Fibrinogen / Q000652:urine*; D006023:Glycoproteins / Q000652:urine*; D006801:Humans; D008297:Male; D008875:Middle Aged; D010455:Peptides / Q000652:urine*; D014552:Urinary Tract Infections / Q000652:urine*
chemical_list: D015415:Biomarkers; D006023:Glycoproteins; D010455:Peptides; D005340:Fibrinogen