6X9B

Structure of proline utilization A with cis-4-hydroxy-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	2018-05-29
Detector	RDI CMOS_8M
Wavelength(s)	1.000
Spacegroup name	P 1 21 1
Unit cell lengths	100.890, 101.920, 126.200
Unit cell angles	90.00, 106.44, 90.00

Refinement procedure

Resolution	48.382 - 1.460
R-factor	0.195
Rwork	0.194
R-free	0.22000
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	5kf6
Data reduction software	XDS
Data scaling software	Aimless (0.7.1)
Phasing software	PHENIX
Refinement software	PHENIX (1.14)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	48.382	48.380	1.480
High resolution limit [Å]	1.460	8.000	1.460
Rmerge	0.062	0.027	1.298
Rmeas	0.071	0.031	1.684
Rpim	0.033	0.013	1.055
Total number of observations		14158	25200
Number of reflections	385095	2609	11043
<I/σ(I)>	12.2	44.4	0.5
Completeness [%]	91.0	96.9	52.8
Redundancy	3.8	5.4	2.3
CC(1/2)	0.998	0.995	0.323

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), cis-4-hydroxy-D-proline (50 mM), and NAD+ (10 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