6X9A

Structure of proline utilization A with trans-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	2018-08-23
Detector	RDI CMOS_8M
Wavelength(s)	1.000
Spacegroup name	P 1 21 1
Unit cell lengths	101.146, 102.005, 126.751
Unit cell angles	90.00, 106.45, 90.00

Refinement procedure

Resolution	45.980 - 1.410
R-factor	0.1764
Rwork	0.175
R-free	0.19880
Structure solution method	FOURIER SYNTHESIS
Starting model (for MR)	5kf6
Data reduction software	XDS
Data scaling software	Aimless (0.7.2)
Phasing software	PHENIX
Refinement software	PHENIX (1.14)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	47.030	47.030	1.430
High resolution limit [Å]	1.410	7.720	1.410
Rmerge	0.060	0.023	1.322
Rmeas	0.071	0.027	1.752
Rpim	0.038	0.014	1.133
Total number of observations	1532777	9684	41774
Number of reflections	456940	2768	18972
<I/σ(I)>	10.7	35	0.5
Completeness [%]	96.5	92	81.3
Redundancy	3.4	3.5	2.2
CC(1/2)	0.998	0.998	0.259

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 trans-4-hydroxy-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