8BS0

Room-temperature structure of Pedobacter heparinus N-acetylglucosamine 2-epimerase at 80 MPa helium gas pressure in a sapphire capillary

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, DESY BEAMLINE P11
Synchrotron site	PETRA III, DESY
Beamline	P11
Temperature [K]	295
Detector technology	PIXEL
Collection date	2020-06-15
Detector	DECTRIS PILATUS3 X CdTe 2M
Wavelength(s)	0.477
Spacegroup name	P 1 21 1
Unit cell lengths	56.350, 93.980, 81.520
Unit cell angles	90.00, 102.55, 90.00

Refinement procedure

Resolution	39.790 - 1.700
R-factor	0.1562
Rwork	0.156
R-free	0.18100
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2gz6
RMSD bond length	0.007
RMSD bond angle	0.792
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHENIX
Refinement software	PHENIX (1.18-3855_9999)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	39.790	1.761
High resolution limit [Å]	1.700	1.700
Rmerge	0.266	1.360
Rmeas	0.282	1.469
Rpim	0.091	0.526
Number of reflections	81812	5880
<I/σ(I)>	5.85	0.99
Completeness [%]	89.8	64.51
Redundancy	8.2	5.3
CC(1/2)	0.985	0.434

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	295	12.6 mg/ml protein in 20 mM sodium phosphate buffer pH 7.5 were mixed (4+2 ul) with 0.1 M sodium cacodylate buffer pH 5.0, 0.35 M sodium acetate and 26% PEG 8000. Crystals grew at room temperature in 5-7 days.