title: Phosphoproteomic analysis reveals regulatory mechanisms at the kidney filtration barrier.
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.
authors: 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.
authors: Markus M
authors: MM
authors: Rinschen
authors: Laboratory of Computational Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland;
authors: Xiongwu
authors: X
authors: Wu
authors: Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases,Institute for Genetics, University of Cologne, Cologne, Germany;
authors: Tim
authors: T
authors: König
authors: Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand;
authors: Trairak
authors: T
authors: Pisitkun
authors: Department of Internal Medicine II, Center for Molecular Medicine.
authors: Henning
authors: H
authors: Hagmann
authors: Department of Internal Medicine II, Center for Molecular Medicine.
authors: Caroline
authors: C
authors: Pahmeyer
authors: Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases.
authors: Tobias
authors: T
authors: Lamkemeyer
authors: Department of Internal Medicine II, Center for Molecular Medicine, Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases.
authors: Priyanka
authors: P
authors: Kohli
authors: Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases.
authors: Nicole
authors: N
authors: Schnell
authors: 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.
authors: Bernhard
authors: B
authors: Schermer
authors: Department of Biology and Biochemistry, University of Houston, Houston, Texas; Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases.
authors: Stuart
authors: S
authors: Dryer
authors: Laboratory of Computational Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland;
authors: Bernard R
authors: BR
authors: Brooks
authors: European Molecular Biology Laboratory-European Bioinformatics Institute, Hinxton, Cambridge, United Kingdom; and.
authors: Pedro
authors: P
authors: Beltrao
authors: Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany.
authors: Marcus
authors: M
authors: Krueger
authors: Department of Internal Medicine II, Center for Molecular Medicine, paul.brinkkoetter@uk-koeln.de thomas.benzing@uk-koeln.de.
authors: Paul T
authors: PT
authors: Brinkkoetter
authors: 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.
authors: Thomas
authors: T
authors: Benzing
mesh_terms: D000818:Animals; D005260:Female; D060730:Glomerular Filtration Barrier / Q000502:physiology*; D047908:Intracellular Signaling Peptides and Proteins / Q000502:physiology; D008565:Membrane Proteins / Q000502:physiology; D051379:Mice; D010750:Phosphoproteins / Q000032:analysis* / Q000502:physiology*; D010766:Phosphorylation; D050199:Podocytes / Q000502:physiology; D040901:Proteomics
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C407223:NPHS2 protein; D010750:Phosphoproteins