title: Automated chip-nanoelectrospray mass spectrometry for glycourinomics in Schindler disease type I.
abstract: In this study an integrative mass spectrometry (MS) approach based on fully automated chip-nanoelectrospray quadrupole time-of-flight was optimized and applied for the discovery and structural characterization of O-glycopeptides in a fraction from the urine of a patient diagnosed with Schindler disease type I. A mixture of O-glycopeptides extracted and purified from an age matched healthy subject served as the control. 49 glycoforms were discovered in the investigated urine fraction from Schindler disease versus only 14 in control urine. Structures with relevant biological significance, previously not described, such as O-fucosylated tetrasaccharides and chains up to pentadecamers O-linked to serine, threonine, or threonine-proline were identified in the pathological urine and characterized by tandem MS (MS/MS). A number of 29 species discovered here, most of which with long chain glycans, were not previously reported as associated to this condition. All glycopeptides were detected in only 1 min analysis time, with a sample consumption situated in the femtomole range.
authors.affiliation: West University of Timisoara, V. Parvan Blvd. 4, RO-300223 Timisoara, Romania; Aurel Vlaicu University of Arad, Revolutiei Blvd. 77, RO-310130 Arad, Romania.
authors.forename: Mirela
authors.initials: M
authors.lastname: Sarbu
authors.affiliation: West University of Timisoara, V. Parvan Blvd. 4, RO-300223 Timisoara, Romania.
authors.forename: Adrian
authors.initials: A
authors.lastname: Robu
authors.affiliation: Westfälische Wilhelms University of Münster, Schlossplatz 2, 48149 Münster, Germany; University of Rijeka, Trg braće Mažuranića 10, 51000 Rijeka, Croatia.
authors.forename: Jasna
authors.initials: J
authors.lastname: Peter-Katalinić
authors.affiliation: West University of Timisoara, V. Parvan Blvd. 4, RO-300223 Timisoara, Romania; National Institute for Research and Development in Electrochemistry and Condensed Matter, Plautius Andronescu Str. 1, RO-300224 Timisoara, Romania. Electronic address: alina.zamfir@uav.ro.
authors.forename: Alina D
authors.initials: AD
authors.lastname: Zamfir
mesh_terms: D001331:Automation; D002240:Carbohydrate Sequence; D002648:Child; D006020:Glycopeptides / Q000737:chemistry / Q000652:urine*; D006801:Humans; D016464:Lysosomal Storage Diseases / Q000652:urine*; D008969:Molecular Sequence Data; D036103:Nanotechnology / Q000379:methods; D019150:Neuroaxonal Dystrophies / Q000652:urine*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D048809:alpha-N-Acetylgalactosaminidase / Q000172:deficiency* / Q000652:urine
chemical_list: D006020:Glycopeptides; D048809:alpha-N-Acetylgalactosaminidase
keywords: Chip-based nanoelectrospray; Lysosomal storage diseases; O-Glycosylated amino acids and peptides; Quadrupole time-of-flight mass spectrometry; Schindler disease type I