9MDD

Crystal Structure of human cyclic GMP-AMP synthase (cGAS) in complex with compound 23; (S)-1-(6,7-dichloro-1-methyl-1,3,4,5-tetrahydro-2H-pyrido[4,3-b]indol-2-yl)-2-methoxyethan-1-one

This is a non-PDB format compatible entry.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS-II BEAMLINE 17-ID-1
Synchrotron site	NSLS-II
Beamline	17-ID-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-10-23
Detector	DECTRIS EIGER X 9M
Wavelength(s)	0.920105
Spacegroup name	P 21 21 2
Unit cell lengths	128.300, 51.710, 60.090
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	37.490 - 1.600
R-factor	0.1953
Rwork	0.194
R-free	0.22020
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	0.968
Data reduction software	DIALS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.19rc6_4061)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.530	1.295
High resolution limit [Å]	1.250	1.250
Rmerge	0.085	0.085
Rmeas		0.091
Rpim		0.033
Number of reflections	73069	110154
<I/σ(I)>	9.44
Completeness [%]	99.5
Redundancy	7.4
CC(1/2)	0.998	0.998

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.