7LJT

Porcine Dihydropyrimidine Dehydrogenase (DPD) soaked with 5-Ethynyluracil (5EU), NADPH - 20 minutes

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	2020-09-27
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.9786
Spacegroup name	P 1 21 1
Unit cell lengths	82.251, 158.861, 162.100
Unit cell angles	90.00, 95.71, 90.00

Refinement procedure

Resolution	60.530 - 1.980
R-factor	0.1848
Rwork	0.183
R-free	0.21900
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1h7x
Data reduction software	xia2
Data scaling software	xia2
Phasing software	PHASER
Refinement software	PHENIX (1.19_4092)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	71.260	71.260	2.010
High resolution limit [Å]	1.980	10.840	1.980
Rmerge	0.138	0.059	0.815
Rpim	0.074	0.033	0.455
Number of reflections	254113	1800	10940
<I/σ(I)>	7.6	19.3	1.6
Completeness [%]	88.6	98.7	77
Redundancy	4.2	4.5	4
CC(1/2)	0.989	0.938	0.615

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	Crystals were obtained by soaking. In each case DPD was crystallized by the vapor diffusion, hanging drop method by mixing 3 uL of 39 uM DPD in 25 mM HEPES, 2 mM DTT, 10% glycerol, pH 7.5 with 3 uL of well solution containing 100 mM NaCl, 2 mM DTT, 18% PEG 6000, pH 4.7. The crystals were then soaked for 20 minutes in 25 mM HEPES, 100 mM NaCl, 2 mM DTT, 100 uM NADPH, 100 uM 5EU, 20% PEG 6000, 20% PEG 400, pH 7.5.