{"corpus_id":85053643,"paper_sha":"55c5ede5581e231c75cc703779075bf4d192d228","doi":"10.1074/JBC.271.37.22552","arxiv_id":null,"pmid":null,"pmcid":null,"mag_id":2081495990,"dblp_id":null,"acl_id":null,"title":"G Protein-coupled Receptor Kinase GRK2 Is a Phospholipid-dependent Enzyme That Can Be Conditionally Activated by G Protein βγ Subunits*","year":1996,"publication_date":"1996-09-13","venue":"Journal of Biological Chemistry","journal":{"name":"The Journal of Biological Chemistry","pages":"22552 - 22562","volume":"271"},"journal_issn":null,"journal_title":null,"publication_types":[],"pubmed_pub_types":null,"s2_fields_of_study":["Biology","Chemistry"],"reference_count":77,"citation_count":102,"influential_citation_count":3,"is_open_access":true,"arxiv_categories":null,"arxiv_license":null,"arxiv_journal_ref":null,"mesh_headings":null,"chemicals":null,"comments_corrections":null,"source_flags":1,"s2_open_access_pdf_url":"http://www.jbc.org/article/S0021925819617898/pdf","s2_open_access_landing_url":"https://www.semanticscholar.org/paper/55c5ede5581e231c75cc703779075bf4d192d228","s2_open_access_license":"CCBY","s2_open_access_status":"HYBRID","pmc_open_access_pdf_url":null,"pmc_open_access_landing_url":null,"pmc_open_access_license":null,"pmc_open_access_status":null,"unpaywall_open_access_pdf_url":null,"unpaywall_open_access_landing_url":null,"unpaywall_open_access_license":null,"unpaywall_open_access_status":null,"abstract":"G protein-coupled receptor kinases (GRKs) mediate agonist-dependent phosphorylation of G protein-coupled receptors (GPRs) and initiate homologous receptor desensitization. Previously, we reported that charged phospholipids directly interacted with the two GRK isoforms, GRK2 and GKR3, via a pleckstrin homology (PH) domain to regulate GRK activity (DebBurman, S. K., Ptasienski, J., Boetticher, E., Lomasney, J. W., Benovic, J. L., and Hosey, M. M. (1995) J. Biol. Chem. 270: 5742-5747). Here, evidence is provided to support the hypothesis that charged phospholipids are required for agonist-dependent phosphorylation of receptors by GRK2. In the absence of charged phospholipids, the purified human m2 muscarinic acetylcholine receptor (hm2mAChR) reconstituted in pure phosphatidylcholine vesicles or in a noninhibitory detergent was not a substrate for GRK2. However, these receptor preparations were stoichiometrically phosphorylated in an agonist-dependent manner upon addition of charged phospholipids. The known ability of G protein βγ subunits to stimulate mAChR phosphorylation also was found to be absolutely dependent on the presence of charged phospholipids, including phosphatidylinositol 4,5-bisphosphate (PIP2). Phospholipids also regulated GRK-mediated phosphorylation of casein, a nonreceptor-soluble substrate. Among lipids tested, lipid inositol phosphates, PIP2 and phosphatidylinositol 4-monophosphate, were found to be the most potent activators of GRK2 and were the only lipids that regulated GRK2 in a complex biphasic manner. At low μM concentrations, PIP2 activated GRK2 via an interaction with the GRK pleckstrin homology domain; however, at high μM concentrations, PIP2 inhibited GRK2, apparently via another mechanism. PIP2-mediated inhibition could be partly relieved by increasing ATP. The results support the hypothesis that GRK2 is a lipid-dependent protein kinase that requires charged phospholipids for enzyme activation, for regulation by Gβγ subunits, and potentially for membrane association.","claims":[{"public_id":"cl_5791931125fa61808d5ffccdb13eccf2","status":"active","text":"At low micromolar concentrations, phosphatidylinositol 4,5-bisphosphate activates GRK2 through its pleckstrin homology domain, whereas at high micromolar concentrations it inhibits GRK2 by a different mechanism that can be partly relieved by increasing ATP.","confidence":0.95,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_5791931125fa61808d5ffccdb13eccf2"},{"public_id":"cl_b7684b215bb00e96bce10d00922a1dfb","status":"active","text":"Charged phospholipids are required for agonist-dependent phosphorylation of receptors by GRK2.","confidence":0.98,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_b7684b215bb00e96bce10d00922a1dfb"},{"public_id":"cl_a026ce3dfa107ed8f783468355c9043f","status":"active","text":"G protein βγ subunit-stimulated mAChR phosphorylation is absolutely dependent on charged phospholipids, including phosphatidylinositol 4,5-bisphosphate.","confidence":0.97,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_a026ce3dfa107ed8f783468355c9043f"},{"public_id":"cl_16416218cf0af7780e9fa67aa324630a","status":"active","text":"Phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol 4-monophosphate are the most potent lipid activators of GRK2 and uniquely regulate it in a complex biphasic manner.","confidence":0.93,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_16416218cf0af7780e9fa67aa324630a"},{"public_id":"cl_31fc5173819ed56200a769069653bfae","status":"active","text":"The purified human m2 muscarinic acetylcholine receptor reconstituted in pure phosphatidylcholine vesicles or in a noninhibitory detergent is not a substrate for GRK2 unless charged phospholipids are added.","confidence":0.96,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_31fc5173819ed56200a769069653bfae"}],"concepts":[{"public_id":"co_022faf784f8370b1e429c947da2dfc71","status":"active","name":"phosphatidylcholine vesicles","description":"Vesicles composed of phosphatidylcholine used as a reconstitution environment 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