6HMK

POLYADPRIBOSYL GLYCOHYDROLASE IN COMPLEX WITH PDD00016690

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I02
Synchrotron site	Diamond
Beamline	I02
Temperature [K]	100
Detector technology	PIXEL
Collection date	2013-07-11
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.979
Spacegroup name	P 21 21 21
Unit cell lengths	67.090, 90.110, 95.500
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	28.770 - 2.060
R-factor	0.1554
Rwork	0.154
R-free	0.18880
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4a0d
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	BUSTER

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	34.830	2.160
High resolution limit [Å]	2.060	2.060
Rmerge	0.092	0.597
Number of reflections	36540
<I/σ(I)>	11.4	3.2
Completeness [%]	100.0	100
Redundancy	5.9	6.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	293	750 nL purified protein at 7.5 mg/mL in 50 mM HEPES, pH 7.0, 150 mM NaCl, 2 mM DTT was mixed with 250 nL of seed stock and 1000 nL of a precipitant consisting of 18-23 % (w/v) PEG-3350, 0.2 M ammonium sulphate, 0.1 M PCTP pH 7.5. Seed stock was prepared using a Seed BeadTM (Hampton Research) from a co-crystal of GS-PARG(448-976 [K617A, Q618A, K619A, E688A, K689A, K690A]) with ADP-ribose, with co-crystallisation mother liquor (19 % (w/v) PEG-3350, 0.2 M ammonium sulphate, 0.1 M PCTP pH 7.5) as the stabilising solution. The final volume of the seed stock was 100 microL.