7KOA

Room Temperature Structure of SARS-CoV-2 Nsp10/16 Methyltransferase in a Complex with Cap-0 and SAM Determined by Pink-Beam Serial Crystallography

Experimental procedure

Experimental method	LAUE
Source type	SYNCHROTRON
Source details	APS BEAMLINE 14-ID-B
Synchrotron site	APS
Beamline	14-ID-B
Temperature [K]	298
Detector technology	CCD
Collection date	2020-09-18
Detector	RAYONIX MX340-HS
Wavelength(s)	1.02-1.18
Spacegroup name	P 31 2 1
Unit cell lengths	170.860, 170.860, 52.797
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	29.590 - 2.400
R-factor	0.2362
Rwork	0.234
R-free	0.27660
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6xkm
RMSD bond length	0.003
RMSD bond angle	1.105
Data reduction software	Precognition
Data scaling software	Epinorm
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.700	2.510
High resolution limit [Å]	2.400	2.400
Rmerge	0.129	0.105
Number of reflections	22795	669
<I/σ(I)>	16.5
Completeness [%]	65.8	15.63
Redundancy	7.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	BATCH MODE	6.5	298	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 polypropylane tube. Crystals were soaked with m7GpppA (0.5 mM) for 10 minutes before data collection.