7QSV
L8-complex forming RubisCO derived from ancestral sequence reconstruction of the last common ancestor of Form I'' and Form I RubisCOs
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 | 2021-08-20 |
Detector | DECTRIS EIGER2 X 16M |
Wavelength(s) | 0.97626 |
Spacegroup name | C 1 2 1 |
Unit cell lengths | 114.007, 203.445, 174.974 |
Unit cell angles | 90.00, 100.43, 90.00 |
Refinement procedure
Resolution | 29.860 - 2.100 |
R-factor | 0.189 |
Rwork | 0.189 |
R-free | 0.22300 |
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] | 29.863 | 29.863 | 2.210 |
High resolution limit [Å] | 2.100 | 6.640 | 2.100 |
Rmerge | 0.044 | 0.911 | |
Rmeas | 0.146 | 0.047 | 0.983 |
Rpim | 0.054 | 0.017 | 0.365 |
Total number of observations | 1613528 | 51877 | 235574 |
Number of reflections | 225493 | 7198 | 32927 |
<I/σ(I)> | 8.9 | 27.2 | 2.1 |
Completeness [%] | 99.2 | 98.6 | 99.3 |
Redundancy | 7.2 | 7.2 | 7.2 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 8 | 288 | Purified enzyme (7.5 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 0.25 mM CABP and 4 mM MgCl2. The enzyme was then mixed in a 1:1 ratio with 40 % v/v Glycerol ethoxylate. Crystals were flash frozen in liquid nitrogen. |