7QSX
Non-obligately L8S8-complex forming RubisCO derived from ancestral sequence reconstruction and rational engineering in L8 complex
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | ESRF BEAMLINE ID23-2 |
Synchrotron site | ESRF |
Beamline | ID23-2 |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2021-07-09 |
Detector | DECTRIS PILATUS3 2M |
Wavelength(s) | 0.873128 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 113.599, 159.197, 127.910 |
Unit cell angles | 90.00, 107.90, 90.00 |
Refinement procedure
Resolution | 29.889 - 2.700 |
R-factor | 0.2189 |
Rwork | 0.218 |
R-free | 0.26030 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 6ura |
Data reduction software | XDS (20210323) |
Data scaling software | SCALA (3.3.22) |
Phasing software | PHENIX (1.18.2_3874) |
Refinement software | PHENIX (1.18.2_3874) |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 29.889 | 29.889 | 2.850 |
High resolution limit [Å] | 2.700 | 8.540 | 2.700 |
Rmerge | 0.092 | 0.821 | |
Rmeas | 0.298 | 0.101 | 0.895 |
Rpim | 0.119 | 0.043 | 0.353 |
Total number of observations | 708705 | 17748 | 107444 |
Number of reflections | 118081 | 3468 | 17291 |
<I/σ(I)> | 5.8 | 11.3 | 2.2 |
Completeness [%] | 99.6 | 91.2 | 100 |
Redundancy | 6 | 5.1 | 6.2 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 6.5 | 288 | Purified enzyme (5 mg/mL) in 25 mM Tricine-NaOH, 75 mM NaCl, pH 8.0 was incubated at 3% (v/v) CO2 in air and 30 degrees C for 1 h in the presence of 0.2 mM CABP and 3.2 mM MgCl2. The enzyme was then mixed in a 1:1 ratio with 0.1 M ammonium sulfate, 0.3 M sodium formate, 0.1 M sodium cacodylate , 3% (w/v) gamma-PGA, and 2% (w/v) PEG 3350, pH 6.5. Before flash freezing the crystals in liquid nitrogen PEG200 was added to the mother liquor as cryoprotectant to a final concentration of 40 % (w/v). |