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Related Experiment Videos

Human urine proteome analysis by three separation approaches.

Wei Sun1, Fuxin Li, Shuzhen Wu

  • 1Proteomics Research Center, National Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, P. R. China.

Proteomics
|November 11, 2005
PubMed
Summary

This study identified 226 urinary proteins, including 12 low molecular mass proteins, advancing the understanding of the human urine proteome. It also explored factors affecting protein identification rates in different proteomics studies.

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Area of Science:

  • Urine proteomics
  • Biomarker discovery
  • Mass spectrometry

Background:

  • The urinary proteome offers insights into organ function, but studies show low overlap.
  • Factors influencing this low overlap, such as methodology, remain unclear.
  • Low molecular mass proteins (<10 kDa) are understudied in urine.

Purpose of the Study:

  • To identify urinary proteins using multiple proteomics approaches.
  • To investigate low molecular mass proteins in urine.
  • To discuss the impact of experimental methods on proteome study overlap.

Main Methods:

  • Collected and pooled male and female urine samples from healthy volunteers.
  • Utilized three protein separation and identification strategies: 1-DE plus 1-D LC/MS/MS, direct 1-D LC/MS/MS, and 2-D LC/MS/MS.

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  • Employed 1-D tricine gels for low molecular mass protein separation and rigorous quality control for mass spectra.

    • Main Results:

    • Identified a total of 226 urinary proteins.
    • Newly identified 171 proteins using proteomics, including 4 male-specific proteins.
    • Characterized 12 low molecular mass proteins and observed post-translational modifications and degradation.

    Conclusions:

    • Established a comprehensive list of urinary proteins, enhancing the human urine proteome database.
    • Highlighted the importance of standardized methods for improving reproducibility in urine proteomics.
    • Provided insights into protein modifications and degradation, crucial for accurate biomarker interpretation.