6VTE

Naegleria gruberi RNA Ligase K170M mutant with AMP and Mn

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-C
Synchrotron site	APS
Beamline	24-ID-C
Temperature [K]	100
Detector technology	PIXEL
Collection date	2015-10-14
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.9792
Spacegroup name	P 32
Unit cell lengths	54.916, 54.916, 102.244
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	27.700 - 2.100
R-factor	0.1955
Rwork	0.189
R-free	0.25310
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	2.140
High resolution limit [Å]	2.100	5.690	2.100
Rmerge	0.093	0.058	0.247
Rmeas	0.116	0.072	0.341
Rpim	0.068	0.042	0.234
Number of reflections	28362	970	483
<I/σ(I)>	10.8
Completeness [%]	86.6	94.8	46.9
Redundancy	2.2	2.7	1.3
CC(1/2)		0.925	0.779

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	NgRnlK170M (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