4BJH

Crystal Structure of the Aquifex Reactor Complex Formed by Dihydroorotase (H180A, H232A) with Dihydroorotate and Aspartate Transcarbamoylase with N-(phosphonacetyl)-L-aspartate (PALA)

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	2009-05-29
Detector	MARRESEARCH MAR300
Spacegroup name	H 3 2
Unit cell lengths	157.152, 157.152, 233.244
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	77.850 - 2.200
R-factor	0.16282
Rwork	0.161
R-free	0.20304
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3d6n
RMSD bond length	0.016
RMSD bond angle	1.558
Data reduction software	MOSFLM
Data scaling software	SCALEPACK
Phasing software	ARP/wARP
Refinement software	REFMAC (5.5.0109)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	77.850	2.320
High resolution limit [Å]	2.200	2.200
Rmerge	0.100	0.760
Number of reflections	53365
<I/σ(I)>	16.2	2.3
Completeness [%]	99.9	100
Redundancy	8.8	8.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5		THE CRYSTALS WERE GROWN AT ROOM TEMPERATURE IN HANGING-DROPS. THE HETERO DODECAMER WAS BUFFER EXCHANGED INTO 10 MM HEPES, 1 MM TCEP PH 7.5, AT A FINAL PROTEIN CONCENTRATION OF 3.0 MG/ML. DHO-ATC SOLUTION (3-6 UL) WAS MIXED WITH 1 UL OF RESERVOIR SOLUTION (30% ETHYLENE GLYCOL) FOLLOWED BY 10% OF THE DROP VOLUME (V/V) OF 100 MM BARIUM CHLORIDE. THE FINAL CRYSTALLIZATION PH WAS 6.3.