Distinct urinary glycoprotein signatures in prostate cancer patients

Rebeca Kawahara, Fabio Ortega, Livia Rosa-Fernandes, Vanessa Guimarães, Daniel Quina, Willian Nahas, Veit Schwämmle, Miguel Srougi, Katia R. M. Leite, Morten Thaysen-Andersen, Martin R. Larsen, Giuseppe Palmisano*

*Corresponding author for this work

Research output: Contribution to journalArticle

12 Citations (Scopus)
30 Downloads (Pure)


Novel biomarkers are needed to complement prostate specific antigen (PSA) in prostate cancer (PCa) diagnostic screening programs. Glycoproteins represent a hitherto largely untapped resource with a great potential as specific and sensitive tumor biomarkers due to their abundance in bodily fluids and their dynamic and cancer-associated glycosylation. However, quantitative glycoproteomics strategies to detect potential glycoprotein cancer markers from complex biospecimen are only just emerging. Here, we describe a glycoproteomics strategy for deep quantitative mapping of N- and O-glycoproteins in urine with a view to investigate the diagnostic value of the glycoproteome to discriminate PCa from benign prostatic hyperplasia (BPH), two conditions that remain difficult to clinically stratify. Total protein extracts were obtained, concentrated and digested from urine of six PCa patients (Gleason score 7) and six BPH patients. The resulting peptide mixtures were TMT-labeled and mixed prior to a multi-faceted sample processing including hydrophilic interaction liquid chromatography (HILIC) and titanium dioxide SPE based enrichment, endo-/exoglycosidase treatment and HILIC-HPLC pre-fractionation. The isolated N- and O-glycopeptides were detected and quantified using high resolution mass spectrometry. We accurately quantified 729 N-glycoproteins spanning 1,310 unique N-glycosylation sites and observed 954 and 965 unique intact N- and O-glycopeptides, respectively, across the two disease conditions. Importantly, a panel of 56 intact N-glycopeptides perfectly discriminated PCa and BPH (ROC: AUC = 1). This study has generated a panel of intact glycopeptides that has a potential for PCa detection.

Original languageEnglish
Pages (from-to)33077-33097
Number of pages21
Issue number69
Publication statusPublished - 4 Sep 2018

Bibliographical note

Copyright the Author(s). Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.


  • prostate cancer
  • urine
  • glycopeptide
  • TMT-labeling
  • glycoproteomics

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    Kawahara, R., Ortega, F., Rosa-Fernandes, L., Guimarães, V., Quina, D., Nahas, W., ... Palmisano, G. (2018). Distinct urinary glycoprotein signatures in prostate cancer patients. Oncotarget, 9(69), 33077-33097. https://doi.org/10.18632/oncotarget.26005