Fig. 5 | Journal of Biomolecular NMR

Fig. 5

From: Efficient production of a functional G protein-coupled receptor in E. coli for structural studies

Fig. 5

Comparison of 1H-15N TROSY spectra of 15N-labeled YY-β1AR derived from E. coli and Sf9 insect cells. Spectra are shown for YY-β1AR binary complexes with the antagonist cyanopindolol and the agonist isoprenaline, as well as the ternary complex with isoprenaline and the G protein-mimicking Nb80. All the experiments were carried out at 304 K, with samples containing ~ 100 μM receptor complexes in 20 mM Tris, 1 mM EDTA, 100 mM NaCl, 37 mM DM, 0.02% NaN3, 5% D2O, pH 7.5. a Superposition of spectra of uniformly 15 N-labeled YY-β1AR obtained from E. coli (blue) and 15 N-valine labeled YY-β1AR obtained from Sf9 cells (red). b Superposition of spectra of uniformly 15N-labeled YY-β1AR obtained from E. coli (blue) and 15N-tyrosine labeled YY-β1AR obtained from Sf9 cells (magenta). Where available, resonances are marked with assignment information. c Crystal structure of β1AR in complex with isoprenaline (green, PDB code 2Y03). Valine residues observed and assigned in panels a and b are shown as red spheres

Back to article page