7L6T

Crystal Structure of SARS-CoV-2 Nsp16/10 Heterodimer in Complex with (m7GpppA2m)pUpUpApApA (Cap-1), S-Adenosyl-L-homocysteine (SAH) and two Magnesium (Mg) ions.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-D
Synchrotron site	APS
Beamline	21-ID-D
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-10-05
Detector	DECTRIS EIGER X 9M
Wavelength(s)	1.12708
Spacegroup name	P 31 2 1
Unit cell lengths	169.351, 169.351, 52.629
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	29.330 - 1.780
R-factor	0.1418
Rwork	0.141
R-free	0.16150
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6w4h
RMSD bond length	0.006
RMSD bond angle	1.330
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	PHASER
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	1.810
High resolution limit [Å]	1.780	1.780
Rmerge	0.090	1.156
Rmeas	0.097	1.269
Rpim	0.036	0.513
Number of reflections	81048	3493
<I/σ(I)>	20.8	2.1
Completeness [%]	96.6	84.2
Redundancy	6.8	5.8
CC(1/2)	0.996	0.560

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	292	Protein: 3.8 mg/ml (nsp10/nsp16 1:1), 0.15M Sodium chloride, 0.01M Tris pH 7.5 , 2mM SAM, 1mM TCEP, 5% Glycerol;Screen: Classics II (B3), 0.5M Magnesium formate, 0.1M HEPES pH 7.5;Soak: 6hours, 0.2mM m7GpppAUUAAA, 5mM SAM, in screen solution;Cryo: 25% Sucrose in screen solution.