5HDI

Structural characterization of CYP144A1, a Mycobacterium tuberculosis cytochrome P450

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I03
Synchrotron site	Diamond
Beamline	I03
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-08-09
Detector	PSI PILATUS 6M
Wavelength(s)	1.00
Spacegroup name	P 21 21 21
Unit cell lengths	58.350, 117.790, 122.090
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	84.770 - 1.540
R-factor	0.14028
Rwork	0.138
R-free	0.19238
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2xkr
RMSD bond length	0.019
RMSD bond angle	1.872
Data reduction software	xia2
Data scaling software	xia2
Phasing software	PHASER
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	88.580	1.580
High resolution limit [Å]	1.540	1.540
Rmerge	0.097	0.807
Number of reflections	116903
<I/σ(I)>	10.4	2.3
Completeness [%]	98.7	97.1
Redundancy	4.4	4.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		277	Crystallography was performed using the sitting drop method using 20 mg/ml CYP144A1. Drops were prepared by the addition of 0.2 microl of CYP144A1-FLV and -TRV proteins to 0.2 microl of mother liquor and by incubating at 4 oC. Following initial crystallogenesis using commercial screens, crystallization conditions were further refined to 0.8 or 1.0 M (NH4)2SO4 with 0.1 M HEPES, pH 7.55, and 25% PEG 3350. Single crystals were flash cooled after addition of 10% PEG 200 as cryoprotectant