8P37
Structure a catalytically inactive mutant of the IMP dehydrogenase related protein GUAB3 from Synechocystis PCC 6803
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | ALBA BEAMLINE XALOC |
| Synchrotron site | ALBA |
| Beamline | XALOC |
| Temperature [K] | 100 |
| Detector technology | PIXEL |
| Collection date | 2022-10-18 |
| Detector | DECTRIS PILATUS 6M |
| Wavelength(s) | 0.979181423454 |
| Spacegroup name | I 4 |
| Unit cell lengths | 112.867, 112.867, 54.119 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 39.910 - 1.219 |
| R-factor | 0.1755 |
| Rwork | 0.175 |
| R-free | 0.18760 |
| Structure solution method | MOLECULAR REPLACEMENT |
| RMSD bond length | 0.013 |
| RMSD bond angle | 1.290 |
| Data reduction software | XDS (20220820) |
| Data scaling software | Aimless (0.7.9) |
| Phasing software | PHASER (2.8.3) |
| Refinement software | BUSTER (2.10.4) |
Data quality characteristics
| Overall | Inner shell | Outer shell | |
| Low resolution limit [Å] | 39.905 | 39.905 | 1.284 |
| High resolution limit [Å] | 1.219 | 3.652 | 1.219 |
| Rmerge | 0.088 | 0.043 | 1.558 |
| Rmeas | 0.091 | 0.045 | 1.628 |
| Rpim | 0.025 | 0.012 | 0.467 |
| Total number of observations | 996816 | 48957 | 46315 |
| Number of reflections | 77746 | 3883 | 3886 |
| <I/σ(I)> | 14.54 | 53.3 | 1.51 |
| Completeness [%] | 94.0 | 99.8 | 80.1 |
| Completeness (spherical) [%] | 76.7 | 99.8 | 26.6 |
| Completeness (ellipsoidal) [%] | 94.0 | 99.8 | 80.1 |
| Redundancy | 12.82 | 12.61 | 11.92 |
| CC(1/2) | 0.999 | 0.999 | 0.680 |
| Anomalous completeness (spherical) | 75.9 | 99.8 | 25.3 |
| Anomalous completeness | 93.8 | 99.8 | 80.0 |
| Anomalous redundancy | 6.5 | 6.6 | 6.2 |
| CC(ano) | -0.339 | -0.444 | -0.031 |
| |DANO|/σ(DANO) | 0.7 | 0.5 | 0.7 |
| Diffraction limits | Principal axes of ellipsoid fitted to diffraction cut-off surface |
| 1.219 Å | 1.000, 1.000, 1.000 |
| 1.219 Å | 0.000, 0.000, 0.000 |
| 1.506 Å | 0.000, 0.000, 0.000 |
| Criteria used in determination of diffraction limits | local <I/sigmaI> ≥ 1.2 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 295 | SynGUAB3-C222S crystals were grown in the presence of IMP and NAD+ in mother liquor from the condition 2-48 of the commercial screen Morpheus (Molecular Dimensions), which consists of an amino acid mixture (0.02M DL-Glutamic acid monohydrate, 0.02M DL-Alanine, 0.02M Glycine, 0.02M DL-Lysine monohydrochloride, and 0.02M DL-Serine, a precipitant mix (25% (v/v) PEG 500 MME and 12.5 % (w/v) PEG 20000), and 0.1M of a buffer system (Tris-base, Bicine) adjusted at pH 8.5. Initial conditions: Buffer: 10 mM TrisHCl, pH 8.0 Protein concentration = 15 mg/mL Substrate concentration: 3 mM NAD + 3 mM IMP |
