9EEH

Crystal structure of E. coli aspartate transcarbamoylase in the R-state complexed with PALA, ATP, GTP, and Mg2+

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	CHESS BEAMLINE 7B2
Synchrotron site	CHESS
Beamline	7B2
Temperature [K]	295
Detector technology	PIXEL
Collection date	2023-06-04
Detector	DECTRIS EIGER2 X 16M
Wavelength(s)	0.9185
Spacegroup name	P 3 2 1
Unit cell lengths	122.436, 122.436, 153.031
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	62.050 - 2.190
R-factor	0.1459
Rwork	0.145
R-free	0.16810
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	0.807
Data reduction software	DIALS (3.18.0)
Data scaling software	DIALS (3.18.0)
Phasing software	PHASER (2.8.3)
Refinement software	PHENIX (1.21.1_5286)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	62.050	2.270
High resolution limit [Å]	2.190	2.190
Rmerge	0.086	0.808
Rpim	0.041	0.378
Number of reflections	67983	6691
<I/σ(I)>	13.9	1.3
Completeness [%]	98.9
Redundancy	5.2
CC(1/2)	0.994	0.705

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	295	Protein solution: 0.016 mM ATCase in 40 mM Tris pH 7.5, 15 mM MgCl2, 1 mM TCEP, 10 mM ATP, 2 mM GTP, and 2 mM N-phosphonacetyl-L-aspartate (PALA) Well solution: Tacsimate pH 7.0 and 9-14% (w/v) PEG 3350 Seed solutions: 10- and 100-fold dilutions were made from a 0.5 mL seed stock prepared by combining two crystallization drops with well solution (21% w/v PEG 3350). Drops: 0.001 mL well solution, 0.001 mL 10- or 100-fold diluted seed solution, 0.002 mL protein solution