Clinical Chemistry
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Clinical Chemistry 53: 1636-1645, 2007. First published July 18, 2007; 10.1373/clinchem.2007.088260
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Right arrow Proteomics and Protein Markers
(Clinical Chemistry. 2007;53:1636-1645.)
© 2007 American Association for Clinical Chemistry, Inc.

Proteomics and Protein Markers

Characterization of Diabetic Nephropathy by Urinary Proteomic Analysis: Identification of a Processed Ubiquitin Form as a Differentially Excreted Protein in Diabetic Nephropathy Patients

Hassan Dihazi1,a, Gerhard A. Müller1, Sandra Lindner1, Markus Meyer2, Abdul R. Asif3, Michael Oellerich3 and Frank Strutz1

1 Department of Nephrology and Rheumatology, Georg-August-University Medical Center, Goettingen, Germany.
2 Indivumed GmbH Center for Cancer Research at the Israelitisches Krankenhaus, Hamburg, Germany.
3 Department of Clinical Chemistry, Georg-August-University Medical Center, Goettingen, Germany.

aAddress correspondence to this author at: Department of Nephrology and Rheumatology, Georg-August University Göttingen, Robert-Koch-Strasse 40, D-37075 Goettingen, Germany. Fax 049-551-3991039; e-mail dihazi{at}med.uni-goettingen.de.

Background: Identification of markers for prediction of the clinical course of diabetic nephropathy remains a major challenge in disease management. We established a proteomics approach for identification of diabetic nephropathy-related biomarkers in urine.

Methods: We used SELDI-TOF mass spectrometry and SAX2 protein arrays to compare protein profiles from urine of 4 defined patient groups. Samples from patients with type 2 diabetes (DM; n = 45) without nephropathy and without microalbuminuria (DM-WNP), patients with DM with macro- or microalbuminuria (DM-NP; n = 38), patients with proteinuria due to nondiabetic renal disease (n = 34), and healthy controls (n = 45) were analyzed. Anionic exchange, reversed-phase fractionation, gel electrophoresis, and mass spectrometry were used to isolate and identify proteins with high discriminatory power.

Results: A protein with m/z 6188 (P <0.0000004) was strongly released in the urine of healthy controls, patients with proteinuria due to nondiabetic disease, and DM-WNP in contrast to DM-NP patients. An m/z 14 766 protein (P <0.00008) was selectively excreted in the urine of DM-NP patients, whereas the protein with m/z 11 774 (P <0.000004) was significantly excreted by patients with proteinuria and DM-NP. The m/z 11 774 and m/z 14 766 mass peaks were identified as ß2-microglobulin and UbA52, a ubiquitin ribosomal fusion protein, respectively. The protein with m/z 6188 was identified as a processed form of ubiquitin.

Conclusion: The release of high amounts of UbA52 in urine of DM-NP patients could serve as a diagnostic marker, whereas the lack of the short form of ubiquitin raises interesting questions about the pathophysiology.




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