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Identification and structural characterization of novel O ‐ and N ‐glycoforms in the urine of a Schindler disease patient by Orbitrap mass spectrometry
Author(s) -
Sarbu Mirela,
Munteanu Cristian V. A.,
Dehelean Liana,
Petrescu Andrei J.,
PeterKatalinic Jasna,
Zamfir Alina D.
Publication year - 2015
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.3616
Subject(s) - orbitrap , chemistry , mass spectrometry , glycan , context (archaeology) , reproducibility , biomarker , chromatography , biomarker discovery , urine , metabolomics , analytical chemistry (journal) , computational biology , proteomics , biochemistry , glycoprotein , gene , paleontology , biology
Schindler disease is an inherited metabolic disorder caused by the deficient activity of α‐N ‐acetylgalactosaminidase enzyme. An accurate diagnosis requires, besides clinical examination, complex and costly biochemical and molecular genetic tests. In the last years, mass spectrometry (MS) based on nanofluidics and high‐resolution instruments has become a successful alternative for disease diagnosis based on the investigation of O ‐glycopeptides in patient urine. A complex mixture of glycoforms extracted from the urine of a 3‐year‐old patient was investigated by Orbitrap MS equipped with Nanospray Flex Ion Source in the negative ion mode. For structural characterization of several molecular species, collision‐induced dissociation MS 2 –MS 3 was carried out using collision energy values within 20–60 eV range. By our approach, 39 novel species associated to this condition were identified, among which O ‐glycopeptides, free O ‐glycans and one structure corresponding to an N ‐glycan never characterized in the context of Schindler disease. The experiments conducted at a resolution of 60 000 allowed the discrimination and identification of a total number of 69 different species with an average mass accuracy of 9.87 ppm, an in‐run reproducibility of almost 100%, an experiment‐to‐experiment and day‐to‐day reproducibility of about 95%. This study brings contributions in the diagnosis of Schindler disease through the elucidation of potential biomarker species in urine. Our multistage MS results completed with 39 new glycoforms the inventory of potential biomarker structures associated to Schindler disease. For the first time, an N ‐glycan was identified and structurally characterized in Schindler patient urine, which opens new research directions in the field. Copyright © 2015 John Wiley & Sons, Ltd.