2I32

Structure of a human ASF1a-HIRA complex and insights into specificity of histone chaperone complex assembly

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X6A
Synchrotron site	NSLS
Beamline	X6A
Temperature [K]	77
Detector technology	CCD
Collection date	2005-04-01
Detector	ADSC QUANTUM 4
Wavelength(s)	0.980
Spacegroup name	P 65 2 2
Unit cell lengths	116.226, 116.226, 167.599
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	25.000 - 2.700
Rwork	0.229
R-free	0.26900
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1roc
RMSD bond length	0.007
RMSD bond angle	1.421
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.590
High resolution limit [Å]	2.500	2.500
Rmerge	0.058	0.641
Number of reflections	43841
<I/σ(I)>	12.8
Completeness [%]	99.9	99.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.6	298	CRYSTALS OF THE HUMAN ASF1AN-HIRA(425-472) COMPLEX WERE GROWN BY HANGING DROP VAPOR DIFFUSION AT ROOM TEMPERATURE AND WERE OBTAINED BY MIXING 2 UL OF A 0.5 MM PROTEIN COMPLEX SOLUTION (IN 20 MM HEPES PH 7.0, 150 MM NACL AND 5 MM BETA-ME) WITH 2 UL OF RESERVOIR SOLUTION CONTAINING 1.44 M NAH2PO4 AND 0.16 M K2HPO4 AT PH 5.6, AND EQUILIBRATING OVER 1.0 ML OF RESERVOIR SOLUTION. CRYSTALS WERE FULLY GROWN WITHIN TWO WEEKS TO A TYPICAL SIZE OF 0.3MMX0.3MMX0.2MM, VAPOR DIFFUSION, HANGING DROP, TEMPERATURE 298.0K