6VTD

Naegleria gruberi RNA ligase R149A 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	2015-11-20
Detector	ADSC QUANTUM 315
Wavelength(s)	0.9792
Spacegroup name	P 41 21 2
Unit cell lengths	88.775, 88.775, 100.926
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	27.050 - 2.400
R-factor	0.2521
Rwork	0.246
R-free	0.30630
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	2.440
High resolution limit [Å]	2.400	6.490	2.400
Rmerge	0.069	0.032	0.800
Rmeas	0.074	0.035	0.867
Rpim	0.028	0.013	0.324
Number of reflections	29868	885	782
<I/σ(I)>	12.4
Completeness [%]	99.1	94.7	100
Redundancy	7.1	6.5	6.8
CC(1/2)		0.999	0.752

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	NgrRnlR149A (8.2 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, 27% PEG6000