2OGY

Asn199Ala Mutant of the 5-methyltetrahydrofolate corrinoid/iron sulfur protein methyltransferase complexed with methyltetrahydrofolate to 2.3 Angstrom resolution

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X25
Synchrotron site	NSLS
Beamline	X25
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2002-05-25
Detector	MARRESEARCH
Spacegroup name	P 21 21 21
Unit cell lengths	49.772, 78.673, 135.857
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	40.190 - 2.300
R-factor	0.18767
Rwork	0.183
R-free	0.23408
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1f6y
RMSD bond length	0.006
RMSD bond angle	0.995
Data reduction software	MOSFLM
Data scaling software	CCP4 ((SCALA))
Phasing software	EPMR
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	46.730	2.360
High resolution limit [Å]	2.300	2.300
Rmerge	0.134	0.385
Number of reflections	24151
<I/σ(I)>	11.1	4.3
Completeness [%]	99.1	99.3
Redundancy	5.4	4.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	298	8-15% PEGMME 5000, 0.05M CALCIUM ACETATE, 20% GLYCEROL, 0.05M HEPES BUFFER; 3-FOLD MOLAR EXCESS OF THE MTHF SUBSTRATE (SCHRICKS LABOLATORIES, JONA, SWITZERLAND). THE SUPERSATURATION OF THE PRECIPITANT SOLUTION REQUIRED DILUTION OF THE PROTEIN-CH3-H4FOLATE COMPLEX BY 50-100-FOLD IN ORDER TO OBTAIN SINGLE CRYSTALS, pH 7.5, VAPOR DIFFUSION, HANGING DROP, temperature 298K