6VTB

Naegleria gruberi RNA ligase K326A 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	2015-10-25
Detector	ADSC QUANTUM 315
Wavelength(s)	0.9792
Spacegroup name	P 21 21 21
Unit cell lengths	53.031, 64.221, 101.888
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	28.600 - 1.547
R-factor	0.1644
Rwork	0.163
R-free	0.19470
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	1.580
High resolution limit [Å]	1.547	4.210	1.547
Rmerge	0.106	0.039	0.914
Rmeas	0.112	0.042	0.975
Rpim	0.035	0.013	0.332
Number of reflections	94052	2778	2370
<I/σ(I)>	10.5
Completeness [%]	96.9	99.2	93.8
Redundancy	9.6	10.3	8.1
CC(1/2)		0.999	0.792

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	NgrRnlK326A (21.4 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