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