7Q03

Ketol-acid reductoisomerase from Methanothermococcus thermolithotrophicus in the close state with NADP and Mg2+

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X10SA
Synchrotron site	SLS
Beamline	X10SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2015-11-30
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.73913
Spacegroup name	I 2 3
Unit cell lengths	129.882, 129.882, 129.882
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	41.070 - 2.100
R-factor	0.1886
Rwork	0.176
R-free	0.18860
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4tsk
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	PHENIX
Refinement software	PHENIX (1.19.2_4158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	45.920	2.210
High resolution limit [Å]	2.100	2.100
Rmerge	0.095	0.744
Rmeas	0.098	0.774
Rpim	0.026	0.212
Number of reflections	21408	3090
<I/σ(I)>	18.3	3.5
Completeness [%]	100.0	100
Redundancy	14.3	13.3
CC(1/2)	0.999	0.468

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	291.15	The reservoir was filled with 90 ul of crystallisation solution: 20% (w/v) PEG 6000, 100 mM Tris/HCl pH 8.0 and 200 mM lithium chloride. The protein was crystallized at 13 mg/ml in the following buffer 25 mM Tris/HCl pH 7.6, 2 mM dithiothreitol, 10% (v/v) glycerol. Drops of 0.7 ul protein with 0.7 ul of crystallisation solution were applied on the shelf. Crystals were soaked in the crystallisation solution supplemented with 30% (v/v) ethylene glycol for few seconds before freezing in liquid nitrogen.