6XKM

Room Temperature Structure of SARS-CoV-2 NSP10/NSP16 Methyltransferase in a Complex with 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]	295
Detector technology	PIXEL
Collection date	2020-06-12
Detector	DECTRIS PILATUS3 X 6M
Wavelength(s)	0.97918
Spacegroup name	P 31 2 1
Unit cell lengths	170.410, 170.410, 52.631
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	49.570 - 2.250
R-factor	0.2068
Rwork	0.206
R-free	0.21260
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6w4h
RMSD bond length	0.005
RMSD bond angle	1.217
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]	49.570	2.290
High resolution limit [Å]	2.250	2.250
Number of reflections	41770	2090
<I/σ(I)>	2.68	0.58
Completeness [%]	100.0	99.7
Redundancy	82.41	10.24
CC(1/2)	0.968	0.449

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	BATCH MODE	6.5	295	Protein: 4.0 mg/ml (NSP10/NSP16 1:1), 0.15 M NaCl, 0.01 M Tris-HCl, 2 mM SAM, 1 mM TCEP, 5% Glycerol, pH 7.5. Precipitation buffer: 0.1 M MES pH 6.5, 0.9 M NaF. Batch crystallization: 100 ul of protein mixed with 100 ul of precipitation buffer in 500 ul polypropylene tube.