2RHI

Crystal structure of the 3-MBT domain from human L3MBTL1 in complex with H1.5K27me2 at 1.66 angstrom

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-C
Synchrotron site	APS
Beamline	24-ID-C
Temperature [K]	100
Detector technology	CCD
Collection date	2007-08-13
Detector	ADSC QUANTUM 315
Wavelength(s)	1.0858
Spacegroup name	P 21 21 2
Unit cell lengths	85.864, 93.318, 58.640
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	20.000 - 1.660
Rwork	0.187
R-free	0.20100
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1oz2
RMSD bond length	0.005
RMSD bond angle	1.410
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	MOLREP
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	1.720
High resolution limit [Å]	1.660	1.660
Rmerge	0.065	0.460
Number of reflections	56401
<I/σ(I)>	31.5	5.1
Completeness [%]	100.0	100
Redundancy	10	9.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	293	L3MBTL1(197-526) (15 mg/ml in 25 mM Tris-HCl, pH 8.0, 1 mM DTT) was pre-incubated with two-fold molar excess of H1.5(23-27)K27me2 peptide. Drops were prepared by mixing 1 l complex solution with 1 l reservoir solution (7% PEG3350, 0.2 M NaAc pH 5.0). Rod-like crystals usually appear overnight and are ready for data collection after two days. Crystals were flash-frozen in liquid nitrogen with the reservoir solution augmented with an additional 30% PEG 400 as cryoprotectant prior to data collectioin., VAPOR DIFFUSION, HANGING DROP, temperature 293K