7MYC

Structure of proline utilization A with the FAD covalently modified by tetrahydrothiophene

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 4.2.2
Synchrotron site	ALS
Beamline	4.2.2
Temperature [K]	100
Detector technology	CMOS
Collection date	2019-10-22
Detector	RDI CMOS_8M
Wavelength(s)	1.00003
Spacegroup name	P 1 21 1
Unit cell lengths	100.932, 101.496, 125.741
Unit cell angles	90.00, 106.54, 90.00

Refinement procedure

Resolution	45.650 - 1.900
R-factor	0.1781
Rwork	0.176
R-free	0.21890
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	5kf6
Data reduction software	XDS
Data scaling software	Aimless (0.7.4)
Phasing software	PHENIX
Refinement software	PHENIX (1.19.2)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	48.380	48.380	1.930
High resolution limit [Å]	1.900	10.410	1.900
Rmerge	0.099	0.026	1.159
Rmeas	0.117	0.031	1.378
Rpim	0.061	0.016	0.737
Total number of observations	677886	4262	32523
Number of reflections	189522	1194	9411
<I/σ(I)>	8.7	28.7	1
Completeness [%]	99.1	96.5	99.9
Redundancy	3.6	3.6	3.5
CC(1/2)	0.996	0.999	0.453

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	286	Protein stock solution: 6 mg/mL PutA with 50 mM tetrahydrothiophene-2-carboxylate in a buffer containing 50 mM Tris (pH 8.0), 50 mM NaCl, 5% (w/v) glycerol, and 0.5 mM Tris(2-caboxyethyl)phosphine. Reservoir solution: 19 % PEG-3350, 0.2 M ammonium sulfate, 0.1 M magnesium chloride, 0.1 M HEPES (pH 8.0), and 0.1 M sodium formate. Cryo-buffer: reservoir supplemented with 15 % PEG-200. Crystals were grown in low-light conditions and then exposed to bright light before flash-cooling to induce the decarboxylation of tetrahydrothiophene-2-carboxylate and covalent modification of the FAD