117E

THE R78K AND D117E ACTIVE SITE VARIANTS OF SACCHAROMYCES CEREVISIAE SOLUBLE INORGANIC PYROPHOSPHATASE: STRUCTURAL STUDIES AND MECHANISTIC IMPLICATIONS

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	EMBL/DESY, HAMBURG BEAMLINE BW7A
Synchrotron site	EMBL/DESY, HAMBURG
Beamline	BW7A
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	1996-09
Detector	MARRESEARCH
Spacegroup name	P 21 21 21
Unit cell lengths	58.400, 102.800, 116.700
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	8.000 - 2.150
R-factor	0.182
Rwork	0.182
R-free	0.24000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1wgj
RMSD bond length	0.007
RMSD bond angle	26.200 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	X-PLOR
Refinement software	X-PLOR (3.851)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.190
High resolution limit [Å]	2.150	2.150
Rmerge	0.086
Total number of observations	228918 *
Number of reflections	36385
<I/σ(I)>	5.7
Completeness [%]	88.9	86
Redundancy	6.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, sitting drop *	6	4 *	17-19% MPD, 25 MM MES, PH 6.0, 1 MM MNCL2, 0.5 MM NA2HPO4, 10 MG/ML PROTEIN

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	10 (mg/ml)
2	1	drop	MES	25 (mM)
3	1	drop		1 (mM)
4	1	drop	sodium hydrogen phosphate	0.5 (mM)
5	1	drop	MPD	17 (%(v/v))
6	1	reservoir	MPD	19 (%(v/v))