7UP7

Crystal structure of C-terminal Domain of MSK1 in complex with covalently bound with literature RSK2 inhibitor indazole cyanoacrylamide compound 26 (soak)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2018-12-13
Detector	RAYONIX MX-300
Wavelength(s)	0.97872
Spacegroup name	P 21 21 21
Unit cell lengths	50.930, 90.520, 135.580
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	47.680 - 2.800
R-factor	0.1877
Rwork	0.181
R-free	0.24440
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	in-house model
RMSD bond length	0.003
RMSD bond angle	0.591
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHENIX
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.870
High resolution limit [Å]	2.800	12.520	2.800
Rmerge	0.086	0.022	0.576
Rmeas	0.095	0.026	0.637
Number of reflections	16052	207	1170
<I/σ(I)>	16.94	42.94	3
Completeness [%]	99.8	94.5	100
Redundancy	5.267	4.111	5.377
CC(1/2)	0.998	0.999	0.853

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	289	Crystals were produced by sitting drop vapor diffusion with an equal volume of the protein, Msk1-C terminal domain (PID7059-1, CID101276) at 5 mg/ml with 1.5mM UCB1590591/BSI106893 in 25mM HEPES pH 7.5, 150mM NaCl, 5% Glycerol, 5mM BME and a crystallization buffer containing 100mM sodium malonate pH 8.0, 100mM Tris pH 8.0, 30% w/v PEG 1000 (tray ID 305760, well F2, CID101276_fine1 screen). Crystals were soaked for 4 days in 2.5ul of reservoir with 1mM UCB1590956/BSI100006.Crystals were cryo protected with 20% EG + compound and vitrified in in liquid N2. Puck ID WJP9-1