Deuteration and selective labeling of alanine methyl groups of beta(2)-adrenergic receptor expressed in a baculovirus-insect cell expression system

作者：Kofuku Yutaka; Yokomizo Tomoki; Imai Shunsuke; Shiraishi Yutaro; Natsume Mei; Itoh Hiroaki; Inoue Masayuki; Nakata Kunio; Igarashi Shunsuke; Yamaguchi Hideyuki; Mizukoshi Toshimi; Suzuki Ei ichiro; Ueda Takumi; Shimada Ichio^*

来源：Journal of Biomolecular NMR, 2018, 71(3): 185-192.

DOI：10.1007/s10858-018-0174-5

• G protein-coupled receptor
• Membrane protein
• Methyl TROSY
• Alanine
• Baculovirus-insect cell expression system

G protein-coupled receptors (GPCRs) exist in equilibrium between multiple conformations, and their populations and exchange rates determine their functions. However, analyses of the conformational dynamics of GPCRs in lipid bilayers are still challenging, because methods for observations of NMR signals of large proteins expressed in a baculovirus-insect cell expression system (BVES) are limited. Here, we report a method to incorporate methyl-(CH3)-C-13-H-1-labeled alanine with > 45% efficiency in highly deuterated proteins expressed in BVES. Application of the method to the NMR observations of beta(2)-adrenergic receptor in micelles and in nanodiscs revealed the ligand-induced conformational differences throughout the transmembrane region of the GPCR.

• 出版日期2018-7