6EPM

Ras guanine nucleotide exchange factor SOS1 (Rem-cdc25) in complex with KRAS(G12C) and fragment screening hit F1

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	BESSY BEAMLINE 14.1
Synchrotron site	BESSY
Beamline	14.1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-08-06
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.918409
Spacegroup name	I 4 2 2
Unit cell lengths	150.223, 150.223, 200.471
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.580 - 2.500
R-factor	0.1813
Rwork	0.180
R-free	0.21088
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	1bkd
RMSD bond length	0.009
RMSD bond angle	1.323
Data reduction software	XDS (VERSION February 3, 2010)
Data scaling software	XDS (VERSION February 3, 2010)
Phasing software	REFMAC
Refinement software	REFMAC (5.8.0158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.600	2.650
High resolution limit [Å]	2.500	2.500
Rmeas	0.118	1.000
Number of reflections	39718	6159
<I/σ(I)>	17.5	2.4
Completeness [%]	99.5	97.1
Redundancy	8.8	8.9
CC(1/2)	0.999	0.771

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	277	Drops made from KRAS SOS1 complex (14.4 mg/ml in 5 mM Tris pH 7.5, 100mM NaCl) and reservoir solution (2.9 to 3.4 M sodium formate, 100 mM MES pH 6.5). Fragment soaked at 25 mM for one day using a 500 mM fragment stock solution in DMSO. Cryo buffer 0.1 M MES pH 6.5, 3.5 M sodium formate, 20 glycerol, 25 mM fragment