Deuteration and selective labeling of alanine methyl groups of β2-adrenergic receptor expressed in a baculovirus-insect cell expression system

• Published in: Journal of biomolecular NMR Vol. 71; no. 3; pp. 185 - 192
• Main Authors: 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
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.07.2018; Springer Nature B.V
• Subjects: Adrenergic receptors; Alanine; Baculovirus; Biochemistry; Biological and Medical Physics; Biophysics; Deuteration; G protein-coupled receptors; Insects; Lipid bilayers; NMR; Nuclear magnetic resonance; Physics; Physics and Astronomy; Proteins; Receptors; Spectroscopy/Spectrometry; G protein-coupled receptor; Methyl TROSY; Membrane protein; Alanine; Baculovirus-insect cell expression system
• ISSN: 0925-2738; 1573-5001
• DOI: 10.1007/s10858-018-0174-5

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- 13 C 1 H 3 -labeled alanine with > 45% efficiency in highly deuterated proteins expressed in BVES. Application of the method to the NMR observations of β 2 -adrenergic receptor in micelles and in nanodiscs revealed the ligand-induced conformational differences throughout the transmembrane region of the GPCR.