24PI

Ancestrally reconstructed acetolactate synthase - Ancestor N1774 with bound ThDP and magnesium.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	AUSTRALIAN SYNCHROTRON BEAMLINE MX2
Synchrotron site	Australian Synchrotron
Beamline	MX2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-11-02
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.953720
Spacegroup name	P 1 21 1
Unit cell lengths	104.383, 126.302, 116.534
Unit cell angles	90.00, 111.39, 90.00

Refinement procedure

Resolution	45.360 - 2.750
R-factor	0.2016
Rwork	0.200
R-free	0.23010
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.003
RMSD bond angle	0.550
Data reduction software	XDS
Data scaling software	Aimless (0.8.2)
Phasing software	PHASER (2.8.3)
Refinement software	PHENIX (2.0_5936)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	48.070	2.810
High resolution limit [Å]	2.750	2.750
Rmerge	0.193	1.279
Rmeas	0.240	1.582
Rpim	0.141	0.915
Number of reflections	71762	4607
<I/σ(I)>	6.9	1.2
Completeness [%]	98.1	97.8
Redundancy	5.4	5.3
CC(1/2)	0.937	0.450

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	293	0.1M Tris.HCl, 30% (w/v) MPEG2000, 1 mM NADH, 10 mM MgCl2