6WMV

Structure of a phosphatidylinositol-phosphate synthase (PIPS) from Mycobacterium kansasii with evidence of substrate binding

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-E
Synchrotron site	APS
Beamline	24-ID-E
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-03-03
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.97918
Spacegroup name	P 1 21 1
Unit cell lengths	78.089, 60.857, 85.417
Unit cell angles	90.00, 90.74, 90.00

Refinement procedure

Resolution	78.083 - 2.142
R-factor	0.2252
Rwork	0.223
R-free	0.26950
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5d91 4o6m
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.13_2998)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	78.083	2.221
High resolution limit [Å]	2.142	2.142
Rmerge	0.073	0.403
Rmeas	0.104	0.571
Rpim	0.073	0.403
Number of reflections	26006	69
<I/σ(I)>	9.78	1.73
Completeness [%]	58.7	1.57
Redundancy	2	2
CC(1/2)	0.995	0.594

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	LIPIDIC CUBIC PHASE	6.1	295	100 mM NaCl, 100 mM Sodium Citrate, pH 6.1, 40 mM MgCl2, 29% PEG 400, 500 uM L-myo-inositol-1-phosphate (prepared in house), 2.5 mM CDP (Sigma) (precipitant). Concentrated protein at 30-35 mg/ml was mixed with monoolein at a 1:1.5 protein to lipid ratio