title: Mapping of the localization of type 1 angiotensin receptor in membrane microdomains using bioluminescence resonance energy transfer-based sensors.
abstract: Initiation and termination of signaling of the type I angiotensin receptor (AT(1)-R) can lead to dynamic changes in its localization in plasma membrane microdomains. Several markers were recently developed to investigate membrane microdomains. Here, we used several YFP-labeled fusion constructs (i.e. raft or non-raft plasma membrane markers) to analyze the agonist-induced changes in compartmentalization of AT(1)-R, including internalization or lateral movement between plasma membrane compartments in response to stimulation using bioluminescence resonance energy transfer measurements. Our data demonstrate that angiotensin II (AngII) stimulus changes the microdomain localization of wild type or mutated (DRY → AAY or TSTS → AAAA) AT(1)-Rs co-expressed with the fluorescent probes in HEK293 cells. The comparison of the trafficking of AT(1)-R upon AngII stimulus with those of [Sar(1),Ile(8)]AngII or [Sar(1),Ile(4),Ile(8)]AngII stimulus revealed different types of changes, depending on the nature of the ligand. The observed changes in receptor compartmentalization of the AT(1)-R are strikingly different from those of 5HT-2C and EGF receptors, which demonstrate the usefulness of the bioluminescence resonance energy transfer-based measurements in the investigation of receptor trafficking in the plasma membrane in living cell experiments.
authors.affiliation: Department of Physiology, Faculty of Medicine, Semmelweis University, Budapest, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Balla
authors.forename: Dániel J
authors.initials: DJ
authors.lastname: Tóth
authors.forename: Eszter
authors.initials: E
authors.lastname: Soltész-Katona
authors.forename: Gyöngyi
authors.initials: G
authors.lastname: Szakadáti
authors.forename: László Sándor
authors.initials: LS
authors.lastname: Erdélyi
authors.forename: Péter
authors.initials: P
authors.lastname: Várnai
authors.forename: László
authors.initials: L
authors.lastname: Hunyady
mesh_terms: D000804:Angiotensin II / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D002460:Cell Line; D004735:Energy Transfer; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D021962:Membrane Microdomains / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D011485:Protein Binding; D021381:Protein Transport; D044140:Receptor, Angiotensin, Type 1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; D044140:Receptor, Angiotensin, Type 1; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D000804:Angiotensin II