8G1H

Ancestral protein AncTh of Phosphomethylpirimidine kinases family

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	LNLS SIRIUS BEAMLINE MANACA
Synchrotron site	LNLS SIRIUS
Beamline	MANACA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-10-07
Detector	DECTRIS PILATUS 2M
Wavelength(s)	0.977180
Spacegroup name	P 61 2 2
Unit cell lengths	59.410, 59.410, 254.936
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	44.010 - 2.700
R-factor	0.2035
Rwork	0.202
R-free	0.22850
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1ub0
RMSD bond length	0.005
RMSD bond angle	0.731
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	MOLREP
Refinement software	PHENIX (1.19.2_4158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	44.010	2.797
High resolution limit [Å]	2.700	2.700
Rmerge	0.193	1.275
Rmeas	0.196	1.293
Rpim	0.033	0.212
Number of reflections	8030	753
<I/σ(I)>	21.62	4.05
Completeness [%]	99.8	99.87
Redundancy	34.1
CC(1/2)	0.999	0.940

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	292	Protein conditions: Protein 26 mg/ml, Tris-HCl 25 mM, NaCl 0.3 M, b-Mercaptoethanol 5 mM, ATP 1 mM, MgCl2 5 mM. Reservoir condition: Ethylene glycols 0.12 M (0.3M Diethylene glycol; 0.3M Triethylene glycol; 0.3M Tetraethylene glycol; 0.3M Pentaethylene glycol), Sodium HEPES/ MOPS pH 7.5, 37.5% precipitants (MPD, PEG 1000, PEG 3350)