7UP5

Crystal structure of C-terminal Domain of MSK1 in complex with covalently bound pyrrolopyrimidine compound 23 (co-crystal)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.2
Synchrotron site	ALS
Beamline	5.0.2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-09-21
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.99999
Spacegroup name	P 21 21 21
Unit cell lengths	51.480, 90.780, 135.970
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.145 - 2.800
R-factor	0.2077
Rwork	0.201
R-free	0.26780
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	in-house model
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHENIX
Refinement software	PHENIX (1.14-3260)

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	0.123	0.026	0.616
Rmeas	0.134	0.029	0.668
Number of reflections	16319	220	1179
<I/σ(I)>	13.83	40.9	3.24
Completeness [%]	100.0	98.2	100
Redundancy	6.495	5.177	6.691
CC(1/2)	0.997	1.000	0.851

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	289	The protein (10.1 mg/ml) was exchanged into 25 mM HEPES pH 7.5, 150 mM NaCl, 5% glycerol, 1 mM DTT using a GE (GE28-9180-04) spin column according to manufacturer's instructions. The protein was then diluted to 0.5 mg/ml in the same buffer and incubated with 150 micromolar UCB170414/BSI108592 for an hour on ice (~10 fold)and concentrated back down to 5 mg/ml. Crystals were produced by sitting drop vapor diffusion with an equal volume of the protein, Msk1-C terminal domain (PID7059-1, CID101276) and a crystallization buffer containing 12.5% PEG 1000, 12.5% PEG 3350, 12.5% MPD: 30mM of each sodium fluoride/iodide/bromide: 100 mM Na HEPES/MOPS pH 8.5 (tray ID 303042, well B12, Morpheus). Crystals were directly vitrified in in liquid N2. Puck ID QRL9-2