7OT0

Human Prolyl-tRNA Synthetase in Complex with L-proline and Compound 4h

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID30B
Synchrotron site	ESRF
Beamline	ID30B
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-12-02
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.976251
Spacegroup name	P 1 21 1
Unit cell lengths	71.163, 93.458, 87.484
Unit cell angles	90.00, 108.61, 90.00

Refinement procedure

Resolution	67.440 - 2.320
R-factor	0.1994
Rwork	0.197
R-free	0.25040
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5vad
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	PHASER
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	67.440	67.440	2.440
High resolution limit [Å]	2.320	7.320	2.320
Rmerge	0.082	0.048	0.968
Rmeas	0.096	0.057	1.131
Rpim	0.049	0.030	0.576
Total number of observations	173295	5079	25709
Number of reflections	46096	1479	6731
<I/σ(I)>	8.5	19.7	1.6
Completeness [%]	97.5	95.8	97.6
Redundancy	3.8	3.4	3.8
CC(1/2)	0.996	0.994	0.732

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	MICROBATCH	7.5	293	The purified protein at 30 mg/mL in 10 mM Tris pH 7, 100 mM NaCl, 2.5 mM 2-mercaptoethanol was incubated with 10 mM L-proline, 2 mM compound and 12% (v/v) DMSO on ice for 1 hr. Crystals were grown in a Terasaki Microbatch plate by mixing 1 uL the pre-mix with 1 uL of reservoir solution containing 0.25-0.4 M SrCl2, 15-20% (v/v) PEG3350 and 100 mM HEPES pH 7.5. The drops were covered with paraffin oil. Crystals were flash frozen in liquid nitrogen directly from the plate.