6VT6

Naegleria gruberi RNA ligase K170A mutant with ATP and Mn

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-E
Synchrotron site	APS
Beamline	24-ID-E
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-03-04
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.9792
Spacegroup name	P 32
Unit cell lengths	54.935, 54.935, 101.647
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	47.575 - 1.969
R-factor	0.1653
Rwork	0.161
R-free	0.21380
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5cot
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	PHENIX
Refinement software	PHENIX (1.16_3549)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.990
High resolution limit [Å]	1.960	5.320	1.960
Rmerge	0.081	0.043	0.395
Rmeas	0.092	0.049	0.469
Rpim	0.045	0.023	0.249
Number of reflections	47215	1216	1131
<I/σ(I)>	7.5
Completeness [%]	99.3	99.7	95.5
Redundancy	4	4.2	3.3
CC(1/2)		0.997	0.781

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	NgRnlK170A (9.8 mg/ml) was adjusted to 5 mM AMP and 2 mM MnCl2 and incubated for 15 min on ice before aliquots of the protein solution (1 ul) were mixed on a coverslip with an equal volume of precipitant solution containing 0.1 M HEPES pH 6.5, 30% PEG6000