Elsevier

Kidney International

Volume 62, Issue 4, October 2002, Pages 1461-1469
Kidney International

Technical Notes
Proteomic analysis of normal human urinary proteins isolated by acetone precipitation or ultracentrifugation

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Proteomic analysis of normal human urinary proteins isolated by acetone precipitation or ultracentrifugation.

Background

Proteomic techniques have recently become available for large-scale protein analysis. The utility of these techniques in identification of urinary proteins is poorly defined. We constructed a proteome map of normal human urine as a reference protein database by using two differential fractionated techniques to isolate the proteins.

Methods

Proteins were isolated from urine obtained from normal human volunteers by acetone precipitation or ultracentrifugation, separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and identified by matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry followed by peptide mass fingerprinting.

Results

A total of 67 protein forms of 47 unique proteins were identified, including transporters, adhesion molecules, complement, chaperones, receptors, enzymes, serpins, cell signaling proteins and matrix proteins. Acetone precipitated more acidic and hydrophilic proteins, whereas ultracentrifugation fractionated more basic, hydrophobic, and membrane proteins. Bioinformatic analysis predicted glycosylation to be the most common explanation for multiple forms of the same protein.

Conclusions

Combining two differential isolation techniques magnified protein identification from human urine. Proteomic analysis of urinary proteins is a promising tool to study renal physiology and pathophysiology and to determine biomarkers of renal disease.

Keywords

protein analysis
transporter
adhesion molecule
chaperone
receptor
post-translational modifications
biomarker
large-scale analysis

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