4LZG

Binary complex of human DNA Polymerase Mu with DNA

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 22-ID
Synchrotron site	APS
Beamline	22-ID
Temperature [K]	93
Detector technology	CCD
Collection date	2012-10-28
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	1.0
Spacegroup name	P 21 21 21
Unit cell lengths	60.003, 68.514, 110.307
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	34.932 - 1.599
R-factor	0.175
Rwork	0.174
R-free	0.20460
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	hPol Mu Loop2 truncation variant in ternary complex with single-nucleotide gapped DNA
RMSD bond length	0.011
RMSD bond angle	1.448
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	PHENIX (1.8_1069)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.630
High resolution limit [Å]	1.599	4.340	1.599
Rmerge		0.042	0.272
Number of reflections	60033
<I/σ(I)>	13.8
Completeness [%]	98.6	99.7	83.1
Redundancy	6.2	6.8	2.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	277	Crystals were grown by mixing 1uL of the protein/DNA complex with 1uL of mother liquor (85mM MES pH 6.5, 85mMCaCl2, 42.5mM NaCl, 8.5% PEG3350, 11% glycerol), using the sitting drop vapor diffusion technique. Crystals were transferred to a cryoprotectant solution containing 0.1M MES pH 6.5, 0.1M CaCl2, 50mM NaCl, 10mM MgCl2, 20% PEG3350, 15% glycerol in three steps. , VAPOR DIFFUSION, SITTING DROP, temperature 277K