9DSQ

Thermotoga maritima threonylcarbamoyl adenylate synthase (TsaC2) in complex with products TC-AMP and pyrophosphate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.2
Synchrotron site	ALS
Beamline	5.0.2
Temperature [K]	80
Detector technology	PIXEL
Collection date	2024-06-08
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.00004
Spacegroup name	P 31 2 1
Unit cell lengths	153.825, 153.825, 86.892
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	44.440 - 1.990
R-factor	0.19828
Rwork	0.197
R-free	0.22250
Structure solution method	FOURIER SYNTHESIS
RMSD bond length	0.004
RMSD bond angle	1.261
Data reduction software	XDS
Data scaling software	pointless (1.12.10)
Phasing software	REFMAC (5.8.0258)
Refinement software	REFMAC (5.8.0430)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	44.440	2.030
High resolution limit [Å]	1.990	1.990
Rmerge	0.271	5.124
Rmeas	0.279	5.263
Rpim	0.064	1.197
Number of reflections	81174	4420
<I/σ(I)>	7.9	0.7
Completeness [%]	100.0	100
Redundancy	18.6
CC(1/2)	0.997	0.322

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	7.4	294.15	Reservoir solution: 500 ul 80 mM sodium cacodylate, pH 7.4, 0.23 M sodium acetate, 13.5% PEG8000, 20% glycerol. Protein: 25 mg/mL TmTsaC2, 50 mM Tris, pH 7.5, 50 mM KCl, 1 mM DTT, 10 mM L-threonine, 30 mM sodium bicarbonate. Crystal soaked in 14% PEG8000, 20% glycerol, 0.2 M sodium acetate, 80 mM sodium cacodylate, pH 7.4, 10 mM L-threonine, 10 mM ATP, 10 mM MgCl2, 30 mM bicarbonate, 0.11 u/ul inorganic pyrophosphatase