7OSY

Human Prolyl-tRNA Synthetase in Complex with L-proline

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE MASSIF-3
Synchrotron site	ESRF
Beamline	MASSIF-3
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-12-02
Detector	DECTRIS EIGER X 4M
Wavelength(s)	0.967700
Spacegroup name	P 1 21 1
Unit cell lengths	69.890, 88.404, 83.745
Unit cell angles	90.00, 110.20, 90.00

Refinement procedure

Resolution	44.200 - 2.230
R-factor	0.2112
Rwork	0.209
R-free	0.26080
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]	78.590	78.590	2.320
High resolution limit [Å]	2.200	6.960	2.200
Rmerge	0.073	0.033	0.864
Rmeas	0.082	0.038	0.967
Rpim	0.037	0.017	0.429
Total number of observations	226553	7016	34398
Number of reflections	47590	1577	6979
<I/σ(I)>	11.3	32.7	1.9
Completeness [%]	98.3	99.3	99.3
Redundancy	4.8	4.4	4.9
CC(1/2)	0.999	0.999	0.713

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 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.