5VOH

Crystal structure of engineered water-forming NADPH oxidase (TPNOX) bound to NADPH. The G159A, D177A, A178R, M179S, P184R mutant of LbNOX.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.2.2
Synchrotron site	ALS
Beamline	8.2.2
Temperature [K]	100
Detector technology	CCD
Collection date	2016-06-04
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.9999
Spacegroup name	P 1 21 1
Unit cell lengths	107.264, 96.527, 119.061
Unit cell angles	90.00, 116.76, 90.00

Refinement procedure

Resolution	47.888 - 2.302
R-factor	0.1942
Rwork	0.192
R-free	0.25510
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5er0
RMSD bond length	0.002
RMSD bond angle	0.525
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	PHENIX (1.10.1_2155)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.340
High resolution limit [Å]	2.300	6.240	2.300
Rmerge	0.102	0.037	0.560
Rmeas	0.109	0.039	0.604
Rpim	0.040	0.014	0.225
Total number of observations	718655
Number of reflections	96175
<I/σ(I)>	6.9
Completeness [%]	100.0	99.9	99.9
Redundancy	7.5	7.6	7.1
CC(1/2)		0.999	0.933

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	294	Crystals were grown in 25 % PEG 3350, 0.1 M Bis-Tris, pH 6.5, 0.2 M (NH4)2SO4 for TPNOX at 21 oC. Crystals were briefly soaked for 1-3 min in the formulation with 15 % (v/v) of ethylene glycol supplemented with 20-30 mM NADPH and were quickly frozen in liquid nitrogen.