2FM7

Evolution of Enzymatic Activity in the Tautomerase Superfamily: Mechanistic and Structural Consequences of the L8R Mutation in 4-Oxalocrotonate Tautomerase

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU300
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2004-02-27
Detector	RIGAKU RAXIS HTC
Wavelength(s)	1.5418
Spacegroup name	P 63
Unit cell lengths	80.863, 80.863, 117.038
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	34.000 - 2.800
R-factor	0.23327
Rwork	0.230
R-free	0.30109
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1BJP with coordinates for OXP removed
RMSD bond length	0.024
RMSD bond angle	2.357
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	MOLREP
Refinement software	REFMAC (5.1.24)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.900
High resolution limit [Å]	2.800	2.800
Rmerge	0.120	0.401
Number of reflections	10209
<I/σ(I)>	5.7
Completeness [%]	99.9	100
Redundancy	7.8	7.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	298	3 microlitres of protein (20 mg/mL solution in 10 mM Tris-Cl, pH 7.0) mixed with an equal volume of reservoir buffer [30% O-(2-aminopropyl)-O-(2-methoxyethyl)polypropylene glycol 500, 100 mM 2-(N-morpholino)ethanesulfonic acid, pH 6.5, and 50 mM CsCl]. The resulting mixture was allowed to equilibrate against 50 microlitres of reservoir solution, VAPOR DIFFUSION, SITTING DROP, temperature 298K