Profiling of the Renal Kinome: a Novel Tool to Identify Protein Kinases Involved in Angiotensin Ii-dependent Hypertensive Renal Damage.
From: Dept. of Pathology, University of Groningen, University Medical Center Groningen, PO Box 30.001, 9700 RB Groningen, The Netherlands. m.h.de.borst@path.umcg.nl
American journal of physiology. Renal physiology
- Publish Date: Jul 2007
- ISSN: 0363-6127
- Volume: 293
- Issue: 1
- Pages: F428-37
- Medium: Print
- Language: English
- Citation (JAMA): de Borst Martin H, Diks Sander H, Bolbrinker Juliane, et al. Profiling of the Renal Kinome: a Novel Tool to Identify Protein Kinases Involved in Angiotensin Ii-dependent Hypertensive Renal Damage.. Am. J. Physiol. Renal Physiol. Jul 2007;293:F428-37
Abstract
Regulation of protein kinase activities is crucial in both physiology and disease, but analysis is hampered by the multitude and complexity of kinase networks. We used novel peptide array chips containing 1,152 known kinase substrate sequences to profile different kinase activities in renal lysates from homozygous Ren2 rats, a model characterized by hypertension and angiotensin II (ANG II)-mediated renal fibrosis, compared with Sprague-Dawley (SD) control rats and Ren2 rats treated with an angiotensin-converting enzyme inhibitor (ACEi). Five-wk-old homozygous Ren2 rats were left untreated or treated with the ACEi ramipril (1 mg.kg(-1).day(-1)) for 4 wk; age-matched SD rats served as controls (n = 5 each). Peptide array chips were incubated with renal cortical lysates in the presence of radioactively labeled ATP. Radioactivity incorporated into the substrate motifs was measured to quantify kinase activity. A number of kinases with modulated activities, which might contribute to renal damage, were validated by Western blotting, immunoprecipitation, and immunohistochemistry. Relevant kinases identified by the peptide array and confirmed using conventional techniques included p38 MAP kinase and PDGF receptor-beta, which were increased in Ren2 and reversed by ACEi. Furthermore, insulin receptor signaling was reduced in Ren2 compared with control rats, and G protein-coupled receptor kinase (GRK) activity decreased in Ren2 + ACEi compared with untreated Ren2 rats. Array-based profiling of tissue kinase activities in ANG II-mediated renal damage provides a powerful tool for identification of relevant kinase pathways in vivo and may lead to novel strategies for therapy.
Mesh Headings (Keywords): Angiotensin II, Angiotensin-Converting Enzyme Inhibitors, Animals, Blood Pressure, Blotting, Western, Gene Expression Regulation, Enzymologic, Genome, Hypertension, Renal, Immunohistochemistry, Immunoprecipitation, Kidney, Male, Phosphorylation, Protein Kinases, Rats, Receptor, Epidermal Growth Factor, Receptor, Insulin, Receptor, Platelet-Derived Growth Factor beta, Receptors, G-Protein-Coupled, Signal Transduction, p38 Mitogen-Activated Protein Kinases
Check for Full Text / PubMed Unique Identifier (PMID): 17429032
This abstract is part of PubMed, a service of the U.S. National Library of Medicine. PubMed includes more than 17 million citations from MEDLINE and other life science journals for biomedical articles. See Copyright and Disclaimers.
Linked medical terms appearing on this page are added by Healia to help readers find more information and are not part of the original PubMed document.
The data herein was last updated on July 8th, 2008 and may not reflect the most current and accurate data available from NLM.