title: Investigation of the fate of type I angiotensin receptor after biased activation.
abstract: Biased agonism on the type I angiotensin receptor (AT1-R) can achieve different outcomes via activation of G protein-dependent and -independent cellular responses. In this study, we investigated whether the biased activation of AT1-R can lead to different regulation and intracellular processing of the receptor. We analyzed β-arrestin binding, endocytosis, and subsequent trafficking steps, such as early and late phases of recycling of AT1-R in human embryonic kidney 293 cells expressing wild-type or biased mutant receptors in response to different ligands. We used Renilla luciferase-tagged receptors and yellow fluorescent protein-tagged β-arrestin2, Rab5, Rab7, and Rab11 proteins in bioluminescence resonance energy transfer measurements to follow the fate of the receptor after stimulation. We found that not only is the signaling of the receptor different upon using selective ligands, but the fate within the cells is also determined by the type of the stimulation. β-arrestin binding and the internalization kinetics of the angiotensin II-stimulated AT1-R differed from those stimulated by the biased agonists. Similarly, angiotensin II-stimulated wild-type AT1-R showed differences compared with a biased mutant AT1-R (DRY/AAY AT1-R) with regards to β-arrestin binding and endocytosis. We found that the differences in the internalization kinetics of the receptor in response to biased agonist stimulation are due to the differences in plasma membrane phosphatidylinositol 4,5-bisphosphate depletion. Moreover, the stability of the β-arrestin binding is a major determinant of the later fate of the internalized AT1-R receptor.
authors: Department of Physiology, Semmelweis University, Faculty of Medicine, Budapest, Hungary (G.S., A.D.T., I.O., L.S.E., P.V., L.H., A.B.), Magyar Tudományos Akadémia-Semmelweis Egyetem Laboratory of Molecular Physiology, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary (L.S.E., P.V., L.H., A.B.); and Section on Molecular Signal Transduction, Program for Developmental Neuroscience, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland (T.B.).
authors: Gyöngyi
authors: G
authors: Szakadáti
authors: Department of Physiology, Semmelweis University, Faculty of Medicine, Budapest, Hungary (G.S., A.D.T., I.O., L.S.E., P.V., L.H., A.B.), Magyar Tudományos Akadémia-Semmelweis Egyetem Laboratory of Molecular Physiology, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary (L.S.E., P.V., L.H., A.B.); and Section on Molecular Signal Transduction, Program for Developmental Neuroscience, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland (T.B.).
authors: András D
authors: AD
authors: Tóth
authors: Department of Physiology, Semmelweis University, Faculty of Medicine, Budapest, Hungary (G.S., A.D.T., I.O., L.S.E., P.V., L.H., A.B.), Magyar Tudományos Akadémia-Semmelweis Egyetem Laboratory of Molecular Physiology, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary (L.S.E., P.V., L.H., A.B.); and Section on Molecular Signal Transduction, Program for Developmental Neuroscience, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland (T.B.).
authors: Ilona
authors: I
authors: Oláh
authors: Department of Physiology, Semmelweis University, Faculty of Medicine, Budapest, Hungary (G.S., A.D.T., I.O., L.S.E., P.V., L.H., A.B.), Magyar Tudományos Akadémia-Semmelweis Egyetem Laboratory of Molecular Physiology, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary (L.S.E., P.V., L.H., A.B.); and Section on Molecular Signal Transduction, Program for Developmental Neuroscience, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland (T.B.).
authors: László Sándor
authors: LS
authors: Erdélyi
authors: Department of Physiology, Semmelweis University, Faculty of Medicine, Budapest, Hungary (G.S., A.D.T., I.O., L.S.E., P.V., L.H., A.B.), Magyar Tudományos Akadémia-Semmelweis Egyetem Laboratory of Molecular Physiology, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary (L.S.E., P.V., L.H., A.B.); and Section on Molecular Signal Transduction, Program for Developmental Neuroscience, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland (T.B.).
authors: Tamas
authors: T
authors: Balla
authors: Department of Physiology, Semmelweis University, Faculty of Medicine, Budapest, Hungary (G.S., A.D.T., I.O., L.S.E., P.V., L.H., A.B.), Magyar Tudományos Akadémia-Semmelweis Egyetem Laboratory of Molecular Physiology, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary (L.S.E., P.V., L.H., A.B.); and Section on Molecular Signal Transduction, Program for Developmental Neuroscience, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland (T.B.).
authors: Péter
authors: P
authors: Várnai
authors: Department of Physiology, Semmelweis University, Faculty of Medicine, Budapest, Hungary (G.S., A.D.T., I.O., L.S.E., P.V., L.H., A.B.), Magyar Tudományos Akadémia-Semmelweis Egyetem Laboratory of Molecular Physiology, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary (L.S.E., P.V., L.H., A.B.); and Section on Molecular Signal Transduction, Program for Developmental Neuroscience, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland (T.B.) Hunyady@eok.sote.hu.
authors: László
authors: L
authors: Hunyady
authors: Department of Physiology, Semmelweis University, Faculty of Medicine, Budapest, Hungary (G.S., A.D.T., I.O., L.S.E., P.V., L.H., A.B.), Magyar Tudományos Akadémia-Semmelweis Egyetem Laboratory of Molecular Physiology, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary (L.S.E., P.V., L.H., A.B.); and Section on Molecular Signal Transduction, Program for Developmental Neuroscience, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland (T.B.).
authors: András
authors: A
authors: Balla
mesh_terms: D019870:1-Phosphatidylinositol 4-Kinase / Q000037:antagonists & inhibitors; D000804:Angiotensin II / Q000494:pharmacology; D019390:Arrestins / Q000235:genetics / Q000378:metabolism; D060807:Bioluminescence Resonance Energy Transfer Techniques; D002462:Cell Membrane / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D006868:Hydrolysis; D008024:Ligands; D049410:Luciferases, Renilla / Q000235:genetics; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism; D054801:Phosphoinositide Phospholipase C / Q000235:genetics / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D044140:Receptor, Angiotensin, Type 1 / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D008024:Ligands; D019269:Phosphatidylinositol 4,5-Diphosphate; D000081082:Phosphoinositide-3 Kinase Inhibitors; D044140:Receptor, Angiotensin, Type 1; D000071557:beta-Arrestins; C485184:enhanced green fluorescent protein; D000804:Angiotensin II; D049452:Green Fluorescent Proteins; D049410:Luciferases, Renilla; D019870:1-Phosphatidylinositol 4-Kinase; D054801:Phosphoinositide Phospholipase C; D043802:GTP-Binding Protein alpha Subunits, Gq-G11