6VT8

Naegleria gruberi RNA ligase E312A mutant with AMP 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	2015-11-20
Detector	ADSC QUANTUM 315
Wavelength(s)	0.9792
Spacegroup name	P 32
Unit cell lengths	54.941, 54.941, 100.602
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	27.410 - 1.998
R-factor	0.1912
Rwork	0.187
R-free	0.23330
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]	30.000	30.000	2.030
High resolution limit [Å]	1.998	5.420	1.998
Rmerge	0.075	0.029	0.580
Rmeas	0.088	0.034	0.688
Rpim	0.046	0.017	0.366
Number of reflections	42536	1138	1195
<I/σ(I)>	17.7
Completeness [%]	99.6	98	100
Redundancy	3.5	3.8	3.4
CC(1/2)		0.999	0.648

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	NgrRnlE312A (9.1 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, 25% PEG6000