7MYA

Structure of proline utilization A with the FAD covalently-modified by 1,3-dithiolane

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	101.176, 101.884, 125.931
Unit cell angles	90.00, 106.49, 90.00

Refinement procedure

Resolution	46.940 - 1.560
R-factor	0.1774
Rwork	0.176
R-free	0.20460
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]	46.940	46.940	1.590
High resolution limit [Å]	1.560	8.540	1.560
Rmerge	0.087	0.026	1.452
Rmeas	0.103	0.031	1.726
Rpim	0.054	0.016	0.923
Total number of observations	1229588	7692	58494
Number of reflections	343612	2127	17103
<I/σ(I)>	8.7	29.8	0.7
Completeness [%]	99.0	96	100
Redundancy	3.6	3.6	3.4
CC(1/2)	0.997	0.998	0.338

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 1,3-dithiolane-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 1,3-dithiolane-2-carboxylate and covalent modification of the FAD