title: An integrated proteomic and peptidomic assessment of the normal human urinome.
abstract: BACKGROUND
  Urine represents an ideal source of clinically relevant biomarkers as it contains a large number of proteins and low molecular weight peptides. The comprehensive characterization of the normal urinary proteome and peptidome can serve as a reference for future biomarker discovery. Proteomic and peptidomic analysis of urine can also provide insight into normal physiology and disease pathology, especially for urogenital diseases.
  
  
  METHODS
  We developed an integrated proteomic and peptidomic analytical protocol in normal urine. We employed ultrafiltration to separate protein and peptide fractions, which were analyzed separately using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) on the Q-Exactive mass spectrometer.
  
  
  RESULTS
  By analyzing six urines from healthy individuals with advanced age, we identified 1754 proteins by proteomic analysis and 4543 endogenous peptides, arising from 566 proteins by peptidomic analysis. Overall, we identified 2091 non-redundant proteins by this integrated approach. In silico protease activity analysis indicated that metalloproteases are predominantly involved in the generation of the endogenous peptide signature. In addition, a number of proteins that were detected in normal urine have previously been implicated in various urological malignancies, including bladder cancer and renal cell carcinoma (RCC).
  
  
  CONCLUSIONS
  We utilized a highly sensitive proteomics approach that enabled us to identify one of the largest sets of protein identifications documented in normal human urine. The raw proteomics and peptidomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD003595.
authors.forename: Ashley
authors.initials: A
authors.lastname: Di Meo
authors.forename: Ihor
authors.initials: I
authors.lastname: Batruch
authors.forename: Arsani G
authors.initials: AG
authors.lastname: Yousef
authors.forename: Maria D
authors.initials: MD
authors.lastname: Pasic
authors.forename: Eleftherios P
authors.initials: EP
authors.lastname: Diamandis
authors.forename: George M
authors.initials: GM
authors.lastname: Yousef
mesh_terms: D000368:Aged; D015415:Biomarkers / Q000652:urine; D002853:Chromatography, Liquid; D005260:Female; D064368:Healthy Volunteers; D006801:Humans; D008297:Male; D008875:Middle Aged; D010455:Peptides / Q000652:urine*; D011506:Proteins / Q000032:analysis*; D040901:Proteomics; D053719:Tandem Mass Spectrometry; D016482:Urinalysis
chemical_list: D015415:Biomarkers; D010455:Peptides; D011506:Proteins