7LOM

Ornithine Aminotransferase (OAT) soaked with its inactivator - (1S,3S)-3-amino-4-(difluoromethylene)cyclohexene-1-carboxylic 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	2020-09-19
Detector	DECTRIS EIGER X 9M
Wavelength(s)	1.1271
Spacegroup name	P 32 2 1
Unit cell lengths	115.610, 115.610, 186.570
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	44.110 - 2.100
R-factor	0.201
Rwork	0.200
R-free	0.23500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1oat
Data reduction software	xia2
Data scaling software	xia2
Phasing software	PHENIX
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	44.110	44.120	2.140
High resolution limit [Å]	2.100	5.700	2.100
Rmeas	0.175	0.071	2.235
Rpim	0.065	0.025	0.825
Total number of observations	579510	31303	29842
Number of reflections	83376	4278	4194
<I/σ(I)>	7.3	22.8	0.9
Completeness [%]	98.4	94.8	99.8
Redundancy	7	7.3	7.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	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 the holoenzyme crystals reached their maximum size within five days, 1 uL of 10 mM 181 was added to the drop with crystals. Within the first three minutes of 181 addition, the hOAT crystals turned their color from yellow to transparent. The crystals were soaked for 44 minutes, transferred into cryoprotective solution (well solution supplemented with 30% glycerol), and then flash-frozen in liquid nitrogen.