8QMQ

Succinic semialdehyde dehydrogenase from E. coli with bound NAD+

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, EMBL c/o DESY BEAMLINE P14 (MX2)
Synchrotron site	PETRA III, EMBL c/o DESY
Beamline	P14 (MX2)
Temperature [K]	100
Detector technology	PIXEL
Collection date	2023-03-23
Detector	DECTRIS EIGER2 X CdTe 16M
Wavelength(s)	0.9763
Spacegroup name	P 21 21 21
Unit cell lengths	92.080, 116.350, 180.070
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	19.920 - 1.700
R-factor	0.1694
Rwork	0.169
R-free	0.18680
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	0.792
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHENIX
Refinement software	PHENIX ((1.20.1_4487: ???))

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	19.920	19.920	1.750
High resolution limit [Å]	1.700	7.610	1.700
Rmerge	0.090	0.036	0.946
Rmeas	0.094	0.038	0.984
Number of reflections	211048	2439	15243
<I/σ(I)>	18.56
Completeness [%]	99.7
Redundancy	13.7
CC(1/2)	0.999	1.000	0.933

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		295	33.6 mg/mL of SAD in 20 mM HEPES-KOH, 50 mM KCl, pH 7.5 was complemented with 5 mM NAD+. The enzyme was then mixed in a 2:1 ratio with 285 mM Bis-Tris-Propane, 8.25 pH , 17 % (w/v) PEG4000. The final drop size was 3 uL. The drop was complemented with PEG200 to a final concentration of 20 % (v/v) before flash freezing the crystals in liquid nitrogen.