6VT9

Naegleria gruberi RNA ligase E227A 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	CCD
Collection date	2015-08-03
Detector	ADSC QUANTUM 315
Wavelength(s)	0.9793
Spacegroup name	P 41 21 2
Unit cell lengths	88.432, 88.432, 105.347
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	27.500 - 2.490
R-factor	0.2272
Rwork	0.219
R-free	0.30550
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 (1.17.1_3660)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	30.000	30.000	2.540
High resolution limit [Å]	2.490	6.760	2.490
Rmerge	0.144	0.060	2.006
Rmeas	0.150	0.064	2.072
Rpim	0.041	0.019	0.516
Number of reflections	27618	857	749
<I/σ(I)>	8.9
Completeness [%]	99.9	98.8	100
Redundancy	14.7	12	16.6
CC(1/2)		0.996	0.645

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	NgrRnlE227A (12 mg/ml) was adjusted to 2 mM ATP and 5 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