7UP8

Crystal structure of C-terminal Domain of MSK1 in complex with covalently bound pyrrolopyrimidine compound 27 (co-crystal)

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	2021-04-08
Detector	RAYONIX MX-300
Wavelength(s)	0.97872
Spacegroup name	P 21 21 21
Unit cell lengths	51.520, 90.910, 136.560
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.200 - 2.900
R-factor	0.1942
Rwork	0.186
R-free	0.26250
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	in-house model
RMSD bond length	0.005
RMSD bond angle	0.780
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHENIX
Refinement software	PHENIX (phenix 1.21)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.980
High resolution limit [Å]	2.900	12.970	2.900
Rmerge	0.103	0.031	0.559
Rmeas	0.119	0.037	0.643
Number of reflections	14778	183	1071
<I/σ(I)>	10.99	24.29	2.7
Completeness [%]	99.6	91.5	100
Redundancy	4.002	3.142	4.103
CC(1/2)	0.995	0.998	0.824

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	289	Plate 320267f8, puck cnl-6-9.Screen:Morpheus_D8_F8_G8_Index_E7. To prepare the ligand complex, PID7059-1 was exchanged to 25 mM HEPES pH 7.5, 150 mM NaCl, 5% glycerol, 1 mM BME (dillution buffer) using a GE (GE28-9180-04) spin column. The protein was diluted to 1 mg/ml in dilution bufffer and 150 ?M UCB1710266 in 100% DMSO_D6 was added. The complex was incubated at 4 C for an hour, then excess ligand was removed by dialysis using a D-tube dialyzer in 250 ml dilution buffer, overnight at 4 ?C. The following day the protein was concentrated to 10.1 mg/ml and setup 200:100 ul protein:well solution. Crystals were flash frozen in 100% well solution