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J Am Soc Nephrol. 2014 Jul;25(7):1509-22. doi: 10.1681/ASN.2013070760. Epub 2014 Feb 7.

Phosphoproteomic analysis reveals regulatory mechanisms at the kidney filtration barrier.

Author information

1
Department of Internal Medicine II, Center for Molecular Medicine, Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, Systems Biology of Ageing Cologne.
2
Laboratory of Computational Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland;
3
Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases,Institute for Genetics, University of Cologne, Cologne, Germany;
4
Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand;
5
Department of Internal Medicine II, Center for Molecular Medicine.
6
Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases.
7
Department of Internal Medicine II, Center for Molecular Medicine, Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases.
8
Department of Biology and Biochemistry, University of Houston, Houston, Texas; Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases.
9
European Molecular Biology Laboratory-European Bioinformatics Institute, Hinxton, Cambridge, United Kingdom; and.
10
Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany.
11
Department of Internal Medicine II, Center for Molecular Medicine, paul.brinkkoetter@uk-koeln.de thomas.benzing@uk-koeln.de.
12
Department of Internal Medicine II, Center for Molecular Medicine, Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, Systems Biology of Ageing Cologne, paul.brinkkoetter@uk-koeln.de thomas.benzing@uk-koeln.de.

Abstract

Diseases of the kidney filtration barrier are a leading cause of ESRD. Most disorders affect the podocytes, polarized cells with a limited capacity for self-renewal that require tightly controlled signaling to maintain their integrity, viability, and function. Here, we provide an atlas of in vivo phosphorylated, glomerulus-expressed proteins, including podocyte-specific gene products, identified in an unbiased tandem mass spectrometry-based approach. We discovered 2449 phosphorylated proteins corresponding to 4079 identified high-confidence phosphorylated residues and performed a systematic bioinformatics analysis of this dataset. We discovered 146 phosphorylation sites on proteins abundantly expressed in podocytes. The prohibitin homology domain of the slit diaphragm protein podocin contained one such site, threonine 234 (T234), located within a phosphorylation motif that is mutated in human genetic forms of proteinuria. The T234 site resides at the interface of podocin dimers. Free energy calculation through molecular dynamic simulations revealed a role for T234 in regulating podocin dimerization. We show that phosphorylation critically regulates formation of high molecular weight complexes and that this may represent a general principle for the assembly of proteins containing prohibitin homology domains.

PMID:
24511133
PMCID:
PMC4073431
DOI:
10.1681/ASN.2013070760
[Indexed for MEDLINE]
Free PMC Article

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