6CUN
Engineered Cytochrome c from Rhodothermus marinus (Rma TDE) bound to carbene intermediate (1-ethoxy-1-oxopropan-2-ylidene)
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | SSRL BEAMLINE BL12-2 |
Synchrotron site | SSRL |
Beamline | BL12-2 |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2017-07-26 |
Detector | DECTRIS PILATUS 6M |
Wavelength(s) | 0.97946 |
Spacegroup name | P 32 2 1 |
Unit cell lengths | 60.343, 60.343, 77.232 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 38.620 - 1.290 |
R-factor | 0.18192 |
Rwork | 0.181 |
R-free | 0.20234 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 6cuk |
RMSD bond length | 0.018 |
RMSD bond angle | 1.814 |
Data reduction software | XDS |
Data scaling software | Aimless |
Phasing software | PHASER |
Refinement software | REFMAC (5.8.0158) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 38.620 | 1.320 |
High resolution limit [Å] | 1.290 | 1.290 |
Number of reflections | 39554 | |
<I/σ(I)> | 21.4 | 2.9 |
Completeness [%] | 100.0 | 100 |
Redundancy | 19.4 | 16.6 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 293 | 1.43-1.48 M (NH4)2SO4 300-400 mM NaSCN 1.5% isopropanol 2:1 ratio of Rma TDE (14 mg/mL in 20mM Tris HCl pH 7.5) to well solution Crystals were soaked in well solution supplemented with 40 mM ethyl 2-diazopropanoate under aerobic conditions for 30 minutes, followed by addition of sodium dithionite (50 mM) and freezing with liquid nitrogen |