6X99

Structure of proline utilization A with D-proline bound in the L-glutamate-gamma-semialdehyde dehydrogenase active site

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	2020-02-11
Detector	RDI CMOS_8M
Wavelength(s)	0.9762
Spacegroup name	P 1 21 1
Unit cell lengths	101.208, 102.316, 127.032
Unit cell angles	90.00, 106.42, 90.00

Refinement procedure

Resolution	46.075 - 1.560
R-factor	0.1757
Rwork	0.174
R-free	0.20370
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 (1.14)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	46.080	46.080	1.590
High resolution limit [Å]	1.560	8.540	1.560
Rmerge	0.082	0.021	1.272
Rmeas	0.097	0.025	1.504
Rpim	0.050	0.013	0.795
Total number of observations	1255558	7850	60757
Number of reflections	350164	2200	17275
<I/σ(I)>	8.1	29.8	0.6
Completeness [%]	99.5	98.3	99
Redundancy	3.6	3.6	3.5
CC(1/2)	0.998	0.999	0.374

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	286	Crystallization experiments were set up with SmPutA (6 mg/mL) and D-proline (50 mM) 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. Crystals were grown using a reservoir solution containing 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. Cryobuffer: reservoir supplemented with 15 % PEG-200