3ERR

Microtubule binding domain from mouse cytoplasmic dynein as a fusion with seryl-tRNA synthetase

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.3.1
Synchrotron site	ALS
Beamline	8.3.1
Temperature [K]	77
Detector technology	CCD
Collection date	2008-04-12
Detector	ADSC QUANTUM 315
Wavelength(s)	1.5
Spacegroup name	H 3
Unit cell lengths	144.302, 144.302, 159.946
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	49.270 - 2.270
R-factor	0.19954
Rwork	0.197
R-free	0.24655
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1sry
RMSD bond length	0.020
RMSD bond angle	1.923
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	REFMAC (5.4.0067)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.270	2.350
High resolution limit [Å]	2.270	2.270
Number of reflections	57296
<I/σ(I)>	18.6	2
Completeness [%]	99.4	94.5
Redundancy	5.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		300	Protein was changed into crystallization buffer (20mM K-HEPES, pH7.5, 10% w/v glycerol, 0.2mM PMSF, 1mM DTT, 4mM Mg-ATP, 0.01% Na-Azide) and concentrated to 18 mg/ml. Crystallization was carried out by setting hanging drops containing 2 ul of protein, (diluted to 13.5mg/ml with 20mM K-Hepes, pH 7.5, 10% glycerol), 0.3 ul 70% glycerol and 1.8 ul of precipitant (20% PEG 4000, 200mM Ammonium sulfate, 100mM Bis-Tris, pH 5.5) over 500ml of the same precipitant solution. Crystals appeared within one day and were of dimensions up to 200 um., VAPOR DIFFUSION, HANGING DROP, temperature 300K