8QMS
Succinic semialdehyde dehydrogenase from E. coli with Q262R substitution and 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 | 91.300, 115.690, 179.610 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 29.460 - 1.900 |
R-factor | 0.1818 |
Rwork | 0.181 |
R-free | 0.21330 |
Structure solution method | MOLECULAR REPLACEMENT |
RMSD bond length | 0.004 |
RMSD bond angle | 0.728 |
Data reduction software | XDS |
Data scaling software | XSCALE |
Phasing software | PHENIX ((1.20.1_4487)) |
Refinement software | PHENIX ((1.20.1_4487: ???)) |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 29.460 | 29.460 | 1.950 |
High resolution limit [Å] | 1.900 | 8.490 | 1.900 |
Rmerge | 0.133 | 0.044 | 0.998 |
Rmeas | 0.138 | 0.046 | 1.036 |
Number of reflections | 149467 | 1809 | 10908 |
<I/σ(I)> | 13.34 | ||
Completeness [%] | 99.5 | ||
Redundancy | 13.8 | ||
CC(1/2) | 0.999 | 0.999 | 0.920 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 8.25 | 295 | 35.0 mg/mL of SAD_Q262R in 20 mM HEPES-KOH, 50 mM KCl, pH 7.5 was supplemented with 5 mM NAD+ and 1 mM TCEP. The enzyme was then mixed in a 1:1 ratio with 285 mM Bis-Tris-Propane, 8.25 pH , 17 % (w/v) PEG4000. The final drop size was 4 uL. The drop was complemented with PEG200 to a final concentration of 16 % (v/v) before flash freezing the crystals in liquid nitrogen. |