Fig. 6 | Journal of Biomolecular NMR

Fig. 6

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

Fig. 6

Comparison of 1H-15N TROSY spectra of 2H,15N-labeled YY-β1AR derived from E. coli and Sf9 insect cells. a 1H-15N TROSY spectra of 17 μM uniformly 2H,15N-labeled TS-β1AR derived from E. coli (blue) and of 90 μM 60% 2H, 90% 15N-labeled TS-β1AR expressed in baculovirus-infected Sf9 insect cells (red). Both samples contained 1 mM of the antagonist alprenolol in 20 mM Tris, 1 mM EDTA, 100 mM NaCl, 37 mM DM, 0.02% NaN3, 5% D2O, pH 7.5. Experiments were carried out at 304 K with total experimental times of 67 h and 10.5 h for the E. coli and Sf9 samples, respectively. b Histogram of intensity ratios (ISf9/IE. coli) of Sf9 and corresponding E. coli 1H-15N TROSY resonances. Intensities ratios of resonances present in both spectra are shown as brown histogram bars. For resonances, which were only detected in the Sf9 sample, ISf9/IE. coli was approximated by the intensity of the Sf9 resonance divided by 3 times the standard deviation of the noise of the E. coli spectrum. These cases then present a lower limit for ISf9/IE. coli and are shown in dark yellow

Back to article page