5D92

Structure of a phosphatidylinositolphosphate (PIP) synthase from Renibacterium Salmoninarum

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-C
Synchrotron site	APS
Beamline	24-ID-C
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-06-09
Detector	PSI PILATUS 6M
Wavelength(s)	0.97910
Spacegroup name	P 1 21 1
Unit cell lengths	89.002, 62.489, 169.759
Unit cell angles	90.00, 99.77, 90.00

Refinement procedure

Resolution	166.970 - 3.620
R-factor	0.2811
Rwork	0.280
R-free	0.29970
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5d91
RMSD bond length	0.016
RMSD bond angle	1.068
Data reduction software	XDS
Data scaling software	Aimless (0.3.11)
Phasing software	PHASER
Refinement software	PHENIX

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	166.970	166.970	3.970
High resolution limit [Å]	3.620	8.870	3.620
Rmerge	0.252	0.042	0.930
Rpim	0.150	0.026	0.552
Total number of observations	78622	5447	18367
<I/σ(I)>	5	19.3	1.4
Completeness [%]	98.9	98.8	97.9
Redundancy	3.7	3.6	3.8
CC(1/2)	0.991	0.998	0.616

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	LIPIDIC CUBIC PHASE		293	30% (v/v) PEG 300, 0.1 M MES pH 6.0, 0.1 M sodium chloride, 0.1 M magnesium chloride (precipitant); Concentrated protein was mixed with molten monoolein in a 1:1.5 (w/w) ratio of protein:lipid using coupled syringes. A Mosquito LCP (TTP Labtech) robot was used to dispense a typical volume of 50-75 nL of protein/lipid mixture onto a 96-well glass sandwich plate, which was covered with 750 nL precipitant solution. Monoolein was doped with 2% CDP-DAG.