3ZZN

5-Mutant (R79W, R151A, E279A, E299A,E313A) Lactate-Dehydrogenase from Thermus thermophillus

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-4
Synchrotron site	ESRF
Beamline	ID14-4
Temperature [K]	100
Detector technology	CCD
Collection date	2009-05-10
Detector	ADSC CCD
Spacegroup name	C 1 2 1
Unit cell lengths	157.123, 59.597, 153.076
Unit cell angles	90.00, 93.37, 90.00

Refinement procedure

Resolution	45.470 - 2.900
R-factor	0.1919
Rwork	0.188
R-free	0.25120
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2v6m
RMSD bond length	0.015
RMSD bond angle	1.704
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX ((PHENIX.REFINE))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	45.530	3.000
High resolution limit [Å]	2.900	2.900
Rmerge	0.103	0.485
Number of reflections	30826
<I/σ(I)>	14.25	3.05
Completeness [%]	96.8	96.8
Redundancy	3.47	3.55

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	9	277	5-MUT TT-LDH WAS CRYSTALLIZED AT 10MG/ML BY THE HANGING-DROP VAPOUR-DIFFUSION METHOD, FOLLOWING THE MIXING OF THE PROTEIN AND THE MOTHER-LIQUOR SOLUTIONS AT A 1:1 RATIO. THE MOTHER LIQUOR SOLUTION WAS 0.1 M HEPES BUFFER PH 9, 18-20% PEG 6000, 2 M LICL2, 500 MICROMOLAR ADP. CRYSTALS GREW WITHIN A WEEK AT 4 DEGREES CELSIUS.