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Study of the Compartmentalization of Type 1 Angiotensin Receptor Using Bioluminescence Resonance Energy Transfer‐based Sensors
Author(s) -
Hunyady László,
Soltész-Katona Eszter,
Várnai Péter,
Balla András
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.lb174
Subject(s) - angiotensin ii , angiotensin ii receptor type 1 , raft , chemistry , lipid raft , receptor , microbiology and biotechnology , biophysics , lipid microdomain , förster resonance energy transfer , membrane , stimulation , medicine , endocrinology , biology , biochemistry , physics , organic chemistry , quantum mechanics , copolymer , fluorescence , polymer
Initiation and termination of signaling of the type I angiotensin receptor (AT1‐R) can lead to dynamic changes in its localization in membrane microdomains. We used YFP‐labeled fusion constructs constructs (i.e. raft‐, or non‐raft plasma membrane markers) to analyze the molecular dynamics of AT1‐R trafficking in response to stimulation in bioluminescence resonance energy transfer (BRET) measurements. Our data demonstrate that angiotensin II (AngII) stimulus changes the microdomain localization of wild type or mutant (DRY/AAY or TSTS/A) AT1‐Rs in HEK293 cells. The comparison of the trafficking of AT1‐R upon AngII stimulus to either [Sar1,Ile8]‐ AngII or [Sar1,Ile4,Ile8]‐AngII stimulus revealed different type of molecular dynamics depending on the nature of the ligand of the AT1‐R. The molecular dynamics of AT1‐R also changes upon cholesterol depletion, which suggests that the mechanism is raft‐dependent. The molecular dynamics of the AT1‐R are strikingly differ from those of 5HT‐2C and EGF receptors, which demonstrate the usefulness of the BRET based measurements in the investigation of receptor trafficking in living cell experiments. These studies should provide valuable information about the distribution and molecular dynamics of AT1‐R upon angiotensin stimulus among membrane microdomains. Supported by Hungarian National Grant OTKA NK‐100883, TÁMOP 4.2.1 B‐09/1/KMR‐2010‐0001.