7TA0

Human Ornithine Aminotransferase (hOAT) soaked with 5-aminovaleric acid

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-D
Synchrotron site	APS
Beamline	21-ID-D
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-07-03
Detector	DECTRIS EIGER X 9M
Wavelength(s)	1.127
Spacegroup name	P 32 2 1
Unit cell lengths	114.906, 114.906, 185.181
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	99.510 - 2.330
R-factor	0.215
Rwork	0.214
R-free	0.23600
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1oat
RMSD bond length	0.002
RMSD bond angle	0.554
Data reduction software	autoPROC
Data scaling software	pointless
Phasing software	PHASER
Refinement software	PHENIX (1.19.2_4158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	99.512	2.366
High resolution limit [Å]	2.326	2.326
Number of reflections	61264	2995
<I/σ(I)>	7
Completeness [%]	99.9	100
Redundancy	15.3
CC(1/2)	0.994	0.330

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.8	293	Once hOAT was purified, it was transferred to a 10 kDa centrifugal filter tube and concentrated to ~6 mg/mL. The holoenzyme crystals were first grown via a hanging drop vapor diffusion method. Each drop contained 2 uL of protein and 2 uL of well solution. The best crystallization condition contained 8% PEG 6000, 100 mM NaCl, 5% glycerol, and 50 mM Tricine pH 7.8. Once holoenzyme crystals reached their maximum size within seven days, 1 uL of 5-aminovaleric acid was added to the drop with crystals. The crystals were soaked for different time periods from 3 to 59 minutes. After soaking, crystals were transferred into a cryoprotective solution (well solution supplemented with 30% glycerol), and then flash-frozen in liquid nitrogen.