6GVS

Engineered glycolyl-CoA reductase comprising 8 mutations with bound NADP+

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID30B
Synchrotron site	ESRF
Beamline	ID30B
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-06-03
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.976251
Spacegroup name	C 1 2 1
Unit cell lengths	362.202, 123.183, 165.096
Unit cell angles	90.00, 109.25, 90.00

Refinement procedure

Resolution	39.133 - 2.579
R-factor	0.1916
Rwork	0.192
R-free	0.20980
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5jfl
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	PHENIX (1.13_2998)
Refinement software	PHENIX

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	39.133	39.133	2.720
High resolution limit [Å]	2.579	8.160	2.580
Rmerge		0.059	0.969
Rmeas	0.150	0.069	1.116
Rpim	0.074	0.035	0.548
Total number of observations	853804	24453	123635
Number of reflections	212592	6805	30472
<I/σ(I)>	7.6	21.1	1.5
Completeness [%]	98.8	96.9	97.4
Redundancy	4	3.6	4.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.4	288	Enzyme (6.5 mg/mL) was mixed with 0.2 M potassium citrate/KOH pH 8.4, 20% (w/v) PEG 3350, and supplemented with NADP+ from a 30 mM stock in a ratio of 2:2:1 (enzyme: crystallization buffer: substrate). The final drop volume was 5 micro liters.