7RIZ

Crystal structure of RPA3624, a beta-propeller lactonase from Rhodopseudomonas palustris, with active-site bound 2-hydroxyquinoline

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-D
Synchrotron site	APS
Beamline	21-ID-D
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-07-25
Detector	DECTRIS EIGER X 9M
Wavelength(s)	1.127230
Spacegroup name	P 32 2 1
Unit cell lengths	48.558, 48.558, 199.122
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	33.190 - 1.710
R-factor	0.1794
Rwork	0.177
R-free	0.20960
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	7ris
RMSD bond length	0.012
RMSD bond angle	1.030
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX (1.19.2_4158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	33.190	1.771
High resolution limit [Å]	1.710	1.710
Rmerge	0.101	1.421
Rmeas	0.104	1.458
Rpim	0.024	0.323
Number of reflections	30590	2983
<I/σ(I)>	17.31	1.65
Completeness [%]	99.3	98.19
Redundancy	19.5	20.2
CC(1/2)	0.999	0.964

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	293	Crystals were grown in a MRC SD2 plate set with a TTP Labtech Mosquito crystallization robot. Protein sample at 9.2mg/mL was inclubated with 5mM 2-hydroxyquinoline at 293K for 1 hour prior to plate setup. The crystal providing the refinement data set was grown by mixing 200 nL protein-2HQ solution with 100 nL reservoir solution, 24% PEG3350, 0.2 M CaCl2, 0.1M bistris pH 6.5. Crystals were cryoprotected by soaking in reservoir solution supplemented to 30% PEG3350 and direct immersion in liquid nitrogen