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Functional Dynamics of Deuterated β 2 ‐Adrenergic Receptor in Lipid Bilayers Revealed by NMR Spectroscopy
Author(s) -
Kofuku Yutaka,
Ueda Takumi,
Okude Junya,
Shiraishi Yutaro,
Kondo Keita,
Mizumura Takuya,
Suzuki Shiho,
Shimada Ichio
Publication year - 2014
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201406603
Subject(s) - chemistry , g protein coupled receptor , nuclear magnetic resonance spectroscopy , lipid bilayer , receptor , biophysics , population , stereochemistry , biochemistry , membrane , biology , demography , sociology
G‐protein‐coupled receptors (GPCRs) exist in conformational equilibrium between active and inactive states, and the former population determines the efficacy of signaling. However, the conformational equilibrium of GPCRs in lipid bilayers is unknown owing to the low sensitivities of their NMR signals. To increase the signal intensities, a deuteration method was developed for GPCRs expressed in an insect cell/baculovirus expression system. The NMR sensitivities of the methionine methyl resonances from the β 2 ‐adrenergic receptor (β 2 AR) in lipid bilayers of reconstituted high‐density lipoprotein (rHDL) increased by approximately 5‐fold upon deuteration. NMR analyses revealed that the exchange rates for the conformational equilibrium of β 2 AR in rHDLs were remarkably different from those measured in detergents. The timescales of GPCR signaling, calculated from the exchange rates, are faster than those of receptor tyrosine kinases and thus enable rapid neurotransmission and sensory perception.