8C8W

Priestia megaterium mupirocin hyper-resistant HIGH motif mutant of type 2 isoleucyl-tRNA synthetase complexed with an isoleucyl-adenylate analogue

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06DA
Synchrotron site	SLS
Beamline	X06DA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-09-28
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.0000356682333689
Spacegroup name	P 43 21 2
Unit cell lengths	107.920, 107.920, 239.360
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	47.310 - 2.290
R-factor	0.1995
Rwork	0.199
R-free	0.23140
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.003
RMSD bond angle	0.510
Data reduction software	XDS (20190806)
Data scaling software	XSCALE (20190806)
Phasing software	PHASER (CCP4Interface 7.0.077)
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.310	2.372
High resolution limit [Å]	2.290	2.290
Rmerge	0.121	2.125
Rmeas	0.126	2.217
Rpim	0.035	0.622
Number of reflections	64159	4825
<I/σ(I)>	16.65	1.2
Completeness [%]	92.6	76.38
Redundancy	13
CC(1/2)	0.999	0.454

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	277	5 mM Ile-AMS and 16.5 mg/ml mut-GVHH-PmIleRS2 were mixed in 1:1 ratio with the well solution containing: 0.2-0.5 M Potassium tartrate, 10-20 % (w/v) PEG3350, 25 mM Hepes-KOH pH=7,5 at 20 oC, 50 mM NaCl. The drop volume of 2 microliters, was equilibrated towards 300 microliters of the well solution at 4 degrees.