7JVL

Structure of the M101A variant of the SidA ornithine hydroxylase complexed with NADP and the FAD in the "out" conformation

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 4.2.2
Synchrotron site	ALS
Beamline	4.2.2
Temperature [K]	100
Detector technology	CMOS
Collection date	2017-07-21
Detector	RDI CMOS_8M
Wavelength(s)	1.0000
Spacegroup name	P 1 21 1
Unit cell lengths	80.229, 153.811, 89.870
Unit cell angles	90.00, 109.28, 90.00

Refinement procedure

Resolution	62.890 - 2.100
R-factor	0.2049
Rwork	0.204
R-free	0.24060
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	6x0i
RMSD bond length	0.008
RMSD bond angle	0.986
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	PHENIX
Refinement software	PHENIX (1.18)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	62.890	62.890	2.140
High resolution limit [Å]	2.100	11.500	2.100
Rmerge	0.082	0.021	0.445
Rmeas	0.098	0.025	0.546
Rpim	0.053	0.013	0.310
Total number of observations	401873	2684	16380
Number of reflections	115533	740	5692
<I/σ(I)>	9.4	30.2	1.7
Completeness [%]	96.6	98.4	95.9
Redundancy	3.5	3.6	2.9
CC(1/2)	0.993	0.999	0.740

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	293	Enzyme stock solution: 8-10 mg/ml SidA (M101A) with 1 mM NADP+ in 25 mM HEPES (pH 7.5) and 100 mM NaCl. Crystallization reservoir: 17-21 % PEG-3350, 0.1 M HEPES (pH 7.5), 0.1 M calcium acetate. Cryo-buffer: 15 % PEG-200, 20 % PEG 3350, 0.1 M HEPES, (pH 7.5), 0.1 M calcium acetate. Drop ratio: 2 enzyme to 1 reservoir