7TZW

The crystal structure of WT CYP199A4 bound to 4-chlorobenzoic acid

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	AUSTRALIAN SYNCHROTRON BEAMLINE MX2
Synchrotron site	Australian Synchrotron
Beamline	MX2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-02-17
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.95373
Spacegroup name	P 1 21 1
Unit cell lengths	41.190, 51.183, 79.027
Unit cell angles	90.00, 92.09, 90.00

Refinement procedure

Resolution	35.967 - 1.462
R-factor	0.1641
Rwork	0.163
R-free	0.18680
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5uvb
RMSD bond length	0.007
RMSD bond angle	0.869
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	PHASER (2.8.2)
Refinement software	PHENIX (1.11.1-2575)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	42.950	42.950	1.490
High resolution limit [Å]	1.460	8.000	1.460
Rmerge	0.171	0.097	1.384
Rmeas	0.186	0.105	1.515
Rpim	0.071	0.041	0.603
Total number of observations	385880	2362	14686
Number of reflections	56716	369	2489
<I/σ(I)>	6.3	19	1
Completeness [%]	99.3	98.5	89.1
Redundancy	6.8	6.4	5.9
CC(1/2)	0.995	0.991	0.515

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.4	289.15	For crystallisation, CYP199A4 was concentrated to approx. 40 mg mL-1 in 50 mM Tris buffer, pH 7.4. Crystals were obtained using the hanging-drop vapour diffusion method at 16 degrees C using 1.2 microlitres of protein with 1.2 microlitres of reservoir solution and equilibrated with 500 microlitres of the same reservoir solution. The crystallisation buffer was 100 mM Bis-Tris buffer (adjusted to pH 5.0-5.75 with acetic acid), 0.2 M magnesium acetate and 20-32% PEG 3350.