6EP8

InhA Y158F mutant in complex with NADH from Mycobacterium tuberculosis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE BM30A
Synchrotron site	ESRF
Beamline	BM30A
Temperature [K]	100
Detector technology	CCD
Collection date	2016-02-14
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.07244
Spacegroup name	P 62 2 2
Unit cell lengths	98.273, 98.273, 139.836
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	42.553 - 1.800
R-factor	0.1468
Rwork	0.145
R-free	0.17510
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4tro
RMSD bond length	0.009
RMSD bond angle	1.129
Data reduction software	XDS
Data scaling software	SCALA (3.3.22)
Phasing software	MOLREP
Refinement software	PHENIX ((1.10.1_2155: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	46.610	1.900
High resolution limit [Å]	1.800	1.800
Rmerge	0.065	1.210
Rmeas	0.067	1.240
Rpim	0.015	0.271
Number of reflections	37644	5382
<I/σ(I)>	35.6	2.9
Completeness [%]	100.0	100
Redundancy	20.5	20.8
CC(1/2)	1.000	0.924

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	287	Thrombin cleaved InhA at 10 mg/ml with 3 to 10 mM of NADH was crystallized in a 24-well crystal plate, Combiclover Junior (Jena Bioscience). The best crystals appeared in drops of 2 ul of enzyme solution, 1 ul of the reservoir solution and 1 ul of distilled water. The reservoir solution contained 100 mM HEPES/NaOH pH 7.5, 50 mM sodium citrate pH 6.5, 7-9 % 2-methyl-2,4-pentanediol. Crystals appeared typically within two to three weeks in this condition.