4ISZ

RNA ligase RtcB in complex with GTP alphaS and Mn(II)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2012-04-16
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97857
Spacegroup name	P 21 21 21
Unit cell lengths	81.108, 138.912, 149.469
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	49.748 - 2.303
R-factor	0.1661
Rwork	0.165
R-free	0.20480
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.017
RMSD bond angle	1.623
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX
Refinement software	PHENIX ((phenix.refine: 1.8_1069))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.340
High resolution limit [Å]	2.300	2.300
Rmerge	0.149
Number of reflections	73420
<I/σ(I)>	12.2
Completeness [%]	98.4	83.1
Redundancy	7.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.5	298	Crystals were grown from one part RtcB solution at 200 micromolar, 1mM MnCl2, 1mM GTPalphaS, mixed with one part reservoir solution, 2.1 M ammonium sulfate, 0.1 M bistris buffer. Crystals were cryoprotected in reservoir solution supplemented with 20% sucrose, 1mM MnCl2 and 1 mM GTPalphaS , pH 5.5, VAPOR DIFFUSION, HANGING DROP, temperature 298K