1O03

Structure of Pentavalent Phosphorous Intermediate of an Enzyme Catalyzed Phosphoryl transfer Reaction observed on cocrystallization with Glucose 6-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	37.311, 54.173, 104.446
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	100.000 * - 1.400
R-factor	0.22
Rwork	0.170
R-free	0.19440 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	SeMeth MAD model of the same protein to 1.7 angstrom
RMSD bond length	0.005
RMSD bond angle	1.190 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	9999.000 *	1.450
High resolution limit [Å]	1.400	1.400
Rmerge	0.073 *	0.130 *
Total number of observations	983130 *
Number of reflections	66302 *
<I/σ(I)>	34.9	15.7
Completeness [%]	98.0 *	88.1 *
Redundancy	14.8	5

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-Glucose6phosphate, 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))