7JPE

Room Temperature Structure of SARS-CoV-2 Nsp10/Nsp16 Methyltransferase in a Complex with m7GpppA Cap-0 and SAM Determined by Fixed-Target Serial Crystallography

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	293
Detector technology	PIXEL
Collection date	2020-08-02
Detector	DECTRIS PILATUS3 X 6M
Wavelength(s)	0.97918
Spacegroup name	P 31 2 1
Unit cell lengths	170.930, 170.930, 52.786
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	49.720 - 2.180
R-factor	0.2179
Rwork	0.217
R-free	0.23700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6wq3
RMSD bond length	0.005
RMSD bond angle	1.351
Data reduction software	DIALS
Data scaling software	DIALS
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0258)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.720	2.220
High resolution limit [Å]	2.180	2.180
Number of reflections	46273	2290
<I/σ(I)>	2.55	0.53
Completeness [%]	100.0	99.7
Redundancy	50.51	11.42
CC(1/2)	0.950	0.398

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	BATCH MODE	6.5	297	Protein: 4.0 mg/ml (NSP10/NSP16 1:1), 0.15 M CaCl, 0.01 M Tris-HCl, 2 mM SAM, 1 mM TCEP, 5% Glycerol, pH 7.5. Precipitation buffer: 0.1M MES pH 6.5, 0.9 M NaF. Batch crystallization: 100 ul of protein mixed with 100 ul of precipitation buffer in 500 ul polypropylane tube. Two days before data collection 1 mM EDTA was added to batch crystallization. Crystal were soaked with m7GpppA (0.5 mM) for 10 minutes before data collection.