4BX9

Human Vps33A in complex with a fragment of human Vps16

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04-1
Synchrotron site	Diamond
Beamline	I04-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2012-05-10
Detector	DECTRIS PILATUS 2M
Spacegroup name	P 43 21 2
Unit cell lengths	128.410, 128.410, 263.360
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	85.840 - 2.600
R-factor	0.17762
Rwork	0.176
R-free	0.21024
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4bx8
RMSD bond length	0.017
RMSD bond angle	1.756
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	87.800	2.720
High resolution limit [Å]	2.600	2.600
Rmerge	0.110	1.310
Number of reflections	68560
<I/σ(I)>	16.8	2.1
Completeness [%]	100.0	100
Redundancy	15.2	14

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6	293	CRYSTALS WERE GROWN IN SITTING DROPS AT 20C. VPS33A:VPS16 COMPLEX (200 NL; A280 = 7.13, APPROX. 8.7 MG/ML) WAS MIXED 1:1 WITH RESERVOIR SOLUTION AND EQUILIBRATED AGAINST 80 UL RESERVOIRS CONTAINING 4% TACSIMATE PH 6.0, 12% PEG 3350 (HAMPTON RESEARCH). CRYSTALS WERE CRYOPROTECTED BY BRIEF IMMERSION IN RESERVOIR SOLUTION SUPPLEMENTED WITH 30% ETHYLENE GLYCOL AND CRYO-COOLED BY PLUNGING INTO LIQUID NITROGEN.