Table 1 Cryo-EM experiment details.
From: Routine sub-2.5 Å cryo-EM structure determination of GPCRs
PAC1R | GLP-1R-TAS | GLP-1R-GLP-1 | ||||
|---|---|---|---|---|---|---|
+VPP | −VPP | +ZLF | −ZLF | +OLA | −OLA | |
Sample preparation | ||||||
Concentration [mg/ml] | 5.1 | 4.6 | 3.7 | |||
Sample volume [µl] | 3 | 3 | 3 | |||
Grid type | Quantifoil R1.2/1.3 Cu/Rh 200 | Quantifoil R1.2/1.3 Cu/Rh 200 | UltrAuFoil R1.2/1.3 Au 300 | |||
Glow discharge time [s] | 30 | 90 | 90 | |||
Glow discharge current [mA] | 10 | 10 | 10 | |||
Blotting chamber temperature [°C] | 4 | 4 | 4 | |||
Blotting chamber humidity [%] | 100 | 100 | 100 | |||
Blot time [s] | 10 | 10 | 10 | |||
Microscopy | ||||||
Voltage | 300 | 300 | 300 | |||
TEM mode | EFTEM Nanoprobe | EFTEM Nanoprobe | EFTEM Nanoprobe | |||
Indicated magnification | 105,000 | 105,000 | 105,000 | |||
C2 aperture [μm] | 50 | 50 | 50 | |||
Spot size | 4 | 4 | 4 | |||
Beam diameter [μm] | 1.55 | 1.41 | 1.45 | |||
Objective lens aperture/phase plate [μm] | VPP | None | 100 | 100 | None | |
Target defocus [μm] | 0.4 | 0.7–1.2 | 0.8–1.5 | 0.6–1.4 | ||
Zero-loss slit width [eV] | 25 | 25 | None | 25 | ||
Pixel size [Å] | 0.83 | 0.83 | 0.83 | |||
Super resolution | No | No | No | |||
Exposure rate [e/pix/s] | 11.75 | 15.3 | 15 | |||
Exposure rate [e/Å2/s] | 17.2 | 22.2 | 21.8 | |||
Exposure time [s] | 3.72 | 3.0 | 3.0 | |||
Total exposure [e/Å2] | 64.0 | 66.5 | 65.4 | |||
Movie frames | 62 | 75 | 75 | |||
Exposure per frame [e/Å2/frame] | 1.03 | 0.89 | 0.87 | |||
