4WVL

Structure-Guided DOT1L Probe Optimization by Label-Free Ligand Displacement

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-D
Synchrotron site	APS
Beamline	21-ID-D
Temperature [K]	100
Detector technology	CCD
Collection date	2013-12-13
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.9792
Spacegroup name	P 65
Unit cell lengths	151.104, 151.104, 53.387
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	43.620 - 2.410
R-factor	0.169
Rwork	0.168
R-free	0.19700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1nw3
RMSD bond length	0.004
RMSD bond angle	0.824
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	MOLREP
Refinement software	PHENIX ((PHENIX.REFINE: 1.9_1692))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.490
High resolution limit [Å]	2.400	2.400
Rmerge	0.085	0.572
Number of reflections	27365
<I/σ(I)>	26.2	5
Completeness [%]	100.0	100
Redundancy	7.5	7.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	4.9	293	A 5-fold excess of 50 mM 5-iodotubercidin (in DMSO) was mixed with protein and crystallized by using the hanging-drop vapor diffusion method at 20 degree in conditions containing 1.8 M (NH4)2SO4, 0.1 M NaAc-pH4.9. Crystals were transferred into stabilization buffer (2.5 M (NH4)2SO4, 0.1 M NaAc-pH4.9, 0.5 M NaCl), incubated with 100 uM inhibitor.