6X25

CRYSTAL STRUCTURE OF INOSITOL POLYPHOSPHATE 1-PHOSPHATASE INPP1 IN COMPLEX GADOLINIUM AFTER ADDITION OF INOSITOL 1,3,4-TRISPHOSPHATE AND LITHIUM AT 3.2 ANGSTROM RESOLUTION

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	296
Detector technology	AREA DETECTOR
Collection date	1994-08-09
Detector	UCSD MARK II
Wavelength(s)	1.542
Spacegroup name	P 41
Unit cell lengths	51.640, 51.640, 143.330
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	10.220 - 3.200
R-factor	0.20161
Rwork	0.199
R-free	0.24551
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	1inp
RMSD bond length	0.007
RMSD bond angle	1.040
Data reduction software	UCSD-system
Data scaling software	UCSD-system
Phasing software	SQUASH
Refinement software	REFMAC (5.8.0258)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	10.220	3.450
High resolution limit [Å]	3.200	3.200
Number of reflections	11374	950
<I/σ(I)>	3.8
Completeness [%]	85.0
Redundancy	2.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.25	293	Tetragonal shaped crystals amenable to diffraction studies were grown by the hanging drop vapor diffusion method on silanized glass covers slips at 20 C and grew with dimensions routinely exceeding 0.1 x 0.1 x 0.5mm. Gd3+, a competitive inhibitor of Mg2+, has a useful anomalous absorption edge at the CuK wavelength and therefore was used in the co-crystallization or soaking conditions. -Mg2+/+Li+/+Ins(1,3,4)P3/+Gd3+ (data=ip7r): Reservoir solution consisted of 10% PEG 8000, 80mM Bis-Tris, pH 5.25, 2mM Gd2(SO4)3, 1mM Ins(1,3,4)P3; 200mM Li2SO4