8CIW

Methylsuccinyl-CoA dehydrogenase from Pseudomonas migulae with bound FAD and (2S)-methylsuccinyl-CoA

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, EMBL c/o DESY BEAMLINE P13 (MX1)
Synchrotron site	PETRA III, EMBL c/o DESY
Beamline	P13 (MX1)
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-09-16
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.9763
Spacegroup name	C 2 2 21
Unit cell lengths	139.805, 169.563, 118.390
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	25.010 - 1.930
R-factor	0.1744
Rwork	0.174
R-free	0.20270
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.009
RMSD bond angle	0.967
Data reduction software	XDS (20190315)
Data scaling software	SCALA (3.3.22)
Phasing software	PHENIX (1.20.1)
Refinement software	PHENIX (1.20.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.010	2.030
High resolution limit [Å]	1.930	1.930
Rmeas	0.124	1.213
Rpim	0.034	0.337
Total number of observations	1387714	191262
Number of reflections	105292	15053
<I/σ(I)>	14.6	2.3
Completeness [%]	99.7
Redundancy	13.2	12.7
CC(1/2)	0.999	0.892

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5	289	Enzyme (20 mg/mL) in 25 mM MOPS pH 7.5 and 0.2 M NaCl was mixed in a 1:1 ratio with crystallization condition (0.2 M ammonium citrate pH 5.0, 20% (w/v) PEG 3350). Soaking and cryoprotection of crystals was performed by adding methylsuccinyl-CoA and PEG200 to final concentrations of 16 mM and 40% (v/v), respectively, before crystals were plunge frozen in liquid nitrogen.