8SKF

Crystal Structure of Betaine aldehyde dehydrogenase (BetB) from Klebsiella aerogenes (Lattice Translocation Disorder)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS-II BEAMLINE 19-ID
Synchrotron site	NSLS-II
Beamline	19-ID
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-02-14
Detector	DECTRIS EIGER2 XE 9M
Wavelength(s)	0.9795
Spacegroup name	C 2 2 21
Unit cell lengths	85.822, 184.823, 163.066
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	81.530 - 1.800
R-factor	0.1758
Rwork	0.174
R-free	0.20740
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.010
RMSD bond angle	0.990
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX ((1.21rc1_4932: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	163.070	1.850
High resolution limit [Å]	1.800	1.800
Rmerge	0.155	1.439
Rmeas	0.161	1.495
Rpim	0.043	0.404
Total number of observations	1639041	118437
Number of reflections	119740	8785
<I/σ(I)>	10.5	1.9
Completeness [%]	100.0
Redundancy	13.7	13.5
CC(1/2)	0.999	0.886

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7	291	JCSG+ C4: 10% (w/v) PEG 6K, 0.10 M HEPES pH 7.0, KlaeA.00020.b.B1.PW39167 at 17 mg/mL. Plate: 13125, well C4 drop 2. Puck: PSL-0714, Cryo: 20% PEG200 + 80% crystallant. 2mM NAD added prior to crystallization. The intensities were corrected for lattice translocation disorder which orginally caused high Rfactors (~25% and 29% for R and Rfree) and residual positive electron density throughout the polypeptide chains. The correction was based on major non-origin Patterson peaks, 1) (0.500, 0.027, 0.500) and its inverse (0.500, -0.027, 0.500) and 2) (0.000, 0.054, 0.000)