6E13

Pseudomonas putida PqqB with a non-physiological zinc at the active site binds the substrate mimic, 5-cysteinyl-3,4-dihydroxyphenylalanine (5-Cys-DOPA), non-specifically but supports the proposed function of the enzyme in pyrroloquinoline quinone biosynthesis.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-B
Synchrotron site	APS
Beamline	23-ID-B
Temperature [K]	295
Detector technology	PIXEL
Collection date	2015-06-14
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.03317
Spacegroup name	P 43 21 2
Unit cell lengths	86.038, 86.038, 107.128
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	45.472 - 2.349
R-factor	0.1917
Rwork	0.189
R-free	0.23560
RMSD bond length	0.007
RMSD bond angle	0.979
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	MOLREP
Refinement software	PHENIX

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.390
High resolution limit [Å]	2.350	6.370	2.350
Rmerge	0.125	0.063	0.927
Rmeas	0.130	0.066	0.967
Rpim	0.037	0.020	0.274
Number of reflections	17434	998	864
<I/σ(I)>	7.5
Completeness [%]	100.0	99.7	100
Redundancy	11.9	10.6	12.2
CC(1/2)		0.997	0.859

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		292	2.5 mg/mL PqqB, 50 mM sodium chloride, 10 mM Tris-HCl, pH 7.5 against reservoir solution for unliganded PqqB (11% PEG4000, 0.2 M sodium chloride, 80 uM zinc chloride, 0.1 M Bis-Tris, pH 6.7)