4PH4

The crystal structure of Human VPS34 in complex with PIK-III

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.2
Synchrotron site	ALS
Beamline	5.0.2
Temperature [K]	100
Detector technology	CCD
Collection date	2009-01-21
Detector	ADSC QUANTUM 315
Wavelength(s)	0.9774
Spacegroup name	P 41 21 2
Unit cell lengths	114.532, 114.532, 146.257
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.340 - 2.800
R-factor	0.1953
Rwork	0.194
R-free	0.22320
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.009
RMSD bond angle	1.070
Data reduction software	XDS
Phasing software	PHASER
Refinement software	BUSTER-TNT (BUSTER 2.11.4)

Data quality characteristics

	Overall
Low resolution limit [Å]	48.750
High resolution limit [Å]	1.980
Number of reflections	29404
<I/σ(I)>	20.9
Completeness [%]	100.0
Redundancy	15.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		303.15	VPS34 protein and PIK-III were mixed and incubated on ice for 1 hr (final PIK-III concentration was 1 mM). Prior to crystallization, the mixture was passed through a 0.2 um filter. The protein:ligand complex was crystallized using the hanging drop vapor diffusion method in Nextal plates: 6 uL of protein solution was mixed with 4 uL of precipitant, which consisted of 20% (w/v) PEG 3350, 100 mM bis-tris propane, and 200 mM Na-K-phosphate. The resulting drop was suspended over a reservoir of 0.3 mL of precipitant and sealed with a screw cap. The crystals grew at 30 degC in approximately 12-24 hr.