1PJT

The structure of the Ser128Ala point-mutant variant of CysG, the multifunctional methyltransferase/dehydrogenase/ferrochelatase for siroheme synthesis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL9-2
Synchrotron site	SSRL
Beamline	BL9-2
Temperature [K]	100
Detector technology	CCD
Collection date	2003-03-10
Detector	ADSC QUANTUM 315
Wavelength(s)	0.979
Spacegroup name	P 21 21 21
Unit cell lengths	59.539, 120.319, 130.036
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	20.000 * - 2.800
R-factor	0.224
Rwork	0.226
R-free	0.29000 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	native CysG PDB entry 1PJQ
RMSD bond length	0.012
RMSD bond angle	1.820 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.900
High resolution limit [Å]	2.800	2.800
Rmerge	0.086 *	0.292 *
Number of reflections	22521
<I/σ(I)>	17.3	4.7
Completeness [%]	93.5	95
Redundancy	4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion *	4.5 *	293	PEG200, phosphate/citrate, sodium chloride, 2-mercaptoethanol, pH 5.0, VAPOR DIFFUSION, HANGING DROP, temperature 293K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	25 (mg/ml)
2	1	reservoir	PEG200	30 (%(v/v))
3	1	reservoir	sodium acetate	200 (mM)	pH4.5
4	1	reservoir		200 (mM)
5	1	reservoir	beta-mercaptoethanol	1 (%(v/v))