9ELX

Crystal Structure of human cyclic GMP-AMP synthase (cGAS) in complex with compound 5; 3-((2-((3-chloro-4-fluorophenyl)amino)-2-oxoethyl)carbamoyl)picolinic acid

This is a non-PDB format compatible entry.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS-II BEAMLINE 17-ID-2
Synchrotron site	NSLS-II
Beamline	17-ID-2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-12-13
Detector	DECTRIS EIGER X 9M
Wavelength(s)	0.979333
Spacegroup name	P 21 21 2
Unit cell lengths	128.246, 51.233, 59.953
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	47.580 - 1.530
R-factor	0.1781
Rwork	0.177
R-free	0.20010
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.014
RMSD bond angle	1.407
Data reduction software	DIALS
Data scaling software	Aimless
Phasing software	PHENIX
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.580	1.585
High resolution limit [Å]	1.530	1.530
Rmerge		0.077
Rmeas		0.084
Rpim		0.031
Number of reflections	60481	60477
<I/σ(I)>	15.17
Completeness [%]	100.0
Redundancy	7.2
CC(1/2)	0.999

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	289	(5mg/ml) HcGAS catalytic domain was mixed with 17/16 bp dsDNA in a molar ratio of 1:1.25 protein:DNA in a buffer containing 20 mM Tris pH 7.5, 150 mM NaCl, 10mM MnCl2, 1mM TCEP, 5mM AMPPNP and 1mM compound. Crystals were obtained with sitting drop vapor diffusion in drops mixed 1:1 with a reservoir of 0.1M Tris pH 8.0, 0.1 M Lithium Sulfate, 20% PEG 4000 and cGAS crystal seeds. Crystals were cryo-protected using reservoir solution supplemented with 25% ethylene glycol and flash-frozen in Liquid N2.