4BX8

Human Vps33A

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I03
Synchrotron site	Diamond
Beamline	I03
Temperature [K]	100
Detector technology	PIXEL
Collection date	2011-05-20
Detector	DECTRIS PILATUS 6M
Spacegroup name	P 1 21 1
Unit cell lengths	71.470, 125.010, 83.280
Unit cell angles	90.00, 115.33, 90.00

Refinement procedure

Resolution	64.610 - 2.400
R-factor	0.19914
Rwork	0.197
R-free	0.24377
Structure solution method	MIRAS
Starting model (for MR)	NONE
RMSD bond length	0.013
RMSD bond angle	1.531
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	autoSHARP
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	64.500	2.490
High resolution limit [Å]	2.400	2.400
Rmerge	0.090	0.770
Number of reflections	49915
<I/σ(I)>	9.5	1.7
Completeness [%]	96.9	97.8
Redundancy	3.5	3.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	CRYSTALS WERE GROWN IN SITTING DROPS AT 20 C. VPS33A-HIS10 (200 NL; 3.2 MG/ML) WAS EQUILIBRATED AGAINST 80 UL RESERVOIRS CONTAINING 14.3-12.8% (W/V) PEG 3350 AND 143-128 MM AMMONIUM ACETATE (HAMPTON RESEARCH). CRYSTALS WERE CRYOPROTECTED BY ADDING RESERVOIR SOLUTION SUPPLEMENTED WITH 30% (V/V) ETHYLENE GLYCOL TO THE MOTHER LIQUOR IMMEDIATELY BEFORE HARVESTING AND CRYO-COOLING BY PLUNGING INTO LIQUID NITROGEN. ANNEALING BY BLOCKING THE CRYOSTREAM FOR 5 S GREATLY ENHANCED THE DIFFRACTION OF VPS33A CRYSTALS.