8BS2

Room-temperature structure of SARS-CoV-2 Main protease at 104 MPa helium gas pressure in a sapphire capillary

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, DESY BEAMLINE P11
Synchrotron site	PETRA III, DESY
Beamline	P11
Temperature [K]	295
Detector technology	PIXEL
Collection date	2021-04-21
Detector	DECTRIS PILATUS3 X CdTe 2M
Wavelength(s)	0.477
Spacegroup name	C 1 2 1
Unit cell lengths	113.580, 54.500, 45.050
Unit cell angles	90.00, 101.25, 90.00

Refinement procedure

Resolution	25.950 - 2.350
R-factor	0.1797
Rwork	0.175
R-free	0.21880
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	7ar5
RMSD bond length	0.003
RMSD bond angle	0.559
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHENIX
Refinement software	PHENIX (1.13-2998_9999)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.950	2.434
High resolution limit [Å]	2.350	2.350
Rmerge	0.285	1.823
Rmeas	0.300	1.963
Rpim	0.088	0.684
Number of reflections	11116	991
<I/σ(I)>	8.4	1.19
Completeness [%]	97.6	86.87
Redundancy	10.2	6.5
CC(1/2)	0.985	0.375

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		291	6.25 mg/ml protein solution in 20 mM HEPES buffer (pH 7.8) containing 1 mM DTT, 1mM EDTA, and 150 mM NaCl was equilibrated against a reservoir solution of 100 mM MIB buffer (2:3:3 molar ratio of malonic acid, imidazole, and boric acid), pH 7.5, containing 25% v/v PEG 1500 and 5% v/v DMSO. To achieve reproducible crystal growth seeding was used. Crystals appeared within a few hours and reached their final size after 2 -3 days.