1O08

Structure of Pentavalent Phosphorous Intermediate of an Enzyme Catalyzed Phosphoryl transfer Reaction observed on cocrystallization with Glucose 1-phosphate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 14-BM-C
Synchrotron site	APS
Beamline	14-BM-C
Temperature [K]	93
Detector technology	CCD
Collection date	2002-02-22
Detector	ADSC QUANTUM 4
Wavelength(s)	0.9
Spacegroup name	P 21 21 21
Unit cell lengths	36.939, 54.297, 104.680
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	20.000 * - 1.200
R-factor	0.1408
Rwork	0.141
R-free	0.17900 *
Structure solution method	AB INITIO
Starting model (for MR)	SeMeth MAD model of the same protein to 1.7 angstrom
RMSD bond length	0.009 *
RMSD bond angle	1.120 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	SHELX
Refinement software	SHELXL-97

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	9999.000 *	1.220
High resolution limit [Å]	1.200	1.200
Rmerge	0.043	0.080 *
Total number of observations	699466 *
Number of reflections	66304 *
<I/σ(I)>	32.7	15.7
Completeness [%]	86.9	54.4
Redundancy	10.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.9	18 *	16% PEG 3350, 0.1M Ammonium Fluoride, 4mM b-D-Glucose-1-phosphate, pH 6.9, VAPOR DIFFUSION, HANGING DROP, temperature 291K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	potassium-HEPES	1 (mM)	pH7.5
2	1	drop		10 (mM)
3	1	drop	dithiothreitol	1 (mM)
4	1	reservoir	ammonium fluoride	0.1 (M)
5	1	reservoir	PEG3350	16 (%(w/v))