8UR1

Crystal structure N-acetylneuraminate lyase (NanA) from Klebsiella aerogenes (pyruvate bound halide free active site)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SEALED TUBE
Source details	BRUKER D8 QUEST
Temperature [K]	100
Detector technology	PIXEL
Collection date	2023-09-26
Detector	Bruker PHOTON III
Wavelength(s)	1.5418
Spacegroup name	P 62 2 2
Unit cell lengths	96.400, 96.400, 205.596
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	46.930 - 2.100
R-factor	0.1843
Rwork	0.183
R-free	0.21420
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.009
RMSD bond angle	0.894
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX ((1.21rc1_5127: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	46.930	2.160
High resolution limit [Å]	2.100	2.100
Rmerge	0.229	1.930
Rmeas	0.232	1.963
Rpim	0.036	0.354
Total number of observations		82554
Number of reflections	33838	2690
<I/σ(I)>	17.9	2
Completeness [%]	100.0
Redundancy	42.6	30.7
CC(1/2)	0.999	0.789

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	291	Morpheus C11: 20%(v/v) Glycerol, 10% w/v PEG 4000, 100 mM Tris/BICINE, pH 8.5, 30 mM NaNO3, 30 mM Na2HPO4 and 30 mM (NH4)2SO4 KlaeA.01563.a.B1.PW39186 at 18.6 mg/mL. 2mM pyruvate added to the protein prior to crystallization. Cryo: direct. This structure has no halides near KPI 165 in the active which resulted in movement of a nearby loop is the previous structures.