5HNK

Crystal structure of T5Fen in complex intact substrate and metal ions.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I24
Synchrotron site	Diamond
Beamline	I24
Temperature [K]	100
Detector technology	PIXEL
Collection date	2011-04-16
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.977
Spacegroup name	P 21 21 21
Unit cell lengths	44.720, 109.940, 127.340
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	42.190 - 2.220
R-factor	0.1851
Rwork	0.183
R-free	0.23440
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1exn
RMSD bond length	0.007
RMSD bond angle	1.066
Data reduction software	xia2
Data scaling software	SCALA (3.3.16)
Phasing software	PHASER
Refinement software	REFMAC (5.8.0135)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	44.720	44.720	2.280
High resolution limit [Å]	2.220	9.930	2.220
Rmerge		0.035	0.654
Rmeas	0.094
Rpim	0.038	0.019	0.405
Total number of observations	182387	2104	6726
Number of reflections	31727
<I/σ(I)>	13.4	28.6	2.2
Completeness [%]	99.4	98.5	94.8
Redundancy	5.7	4.9	3.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.5	290	The concentration of both oligonucleotides was adjusted to 1.1 mM for the duplex molecule by dissolving each one in 10 mM MES pH 6.5 and 50 mM KCl. They were annealed by heating to 367K for 10 minutes and allowed to cool to room temperature. Crystals with oligonucleotide 5ov4 were grown at 290K with the D153K variant of T5Fen. The resulting T5FenD153K:5ov4 structure was determined from crystals grown in 0.2 M MgCl2, 0.1 M Bis-Tris buffer pH 5.5, 25% w/v PEG 3350.