5VG1

Neutron crystallographic structure of a Jonesia denitrificans lytic polysaccharide monooxygenase

Experimental procedure

Experimental method	LAUE
Source type	SPALLATION SOURCE
Source details	ORNL Spallation Neutron Source BEAMLINE MANDI
Synchrotron site	ORNL Spallation Neutron Source
Beamline	MANDI
Temperature [K]	295
Detector technology	AREA DETECTOR
Collection date	2015-05-21
Detector	ORNL ANGER CAMERA
Wavelength(s)	2-4
Spacegroup name	P 21 21 21
Unit cell lengths	32.500, 76.400, 122.100
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	12.000 - 2.100
R-factor	0.191
Rwork	0.187
R-free	0.26500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5vg0
RMSD bond length	0.002
RMSD bond angle	0.560
Data reduction software	Mantid
Phasing software	PHENIX
Refinement software	PHENIX ((1.10.1_2155))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	12.000	2.170
High resolution limit [Å]	2.100	2.100
Number of reflections	13989
<I/σ(I)>	9.4	2.7
Completeness [%]	76.0	59.4
Redundancy	4.5	2.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	Purified enzyme was incubated with a threefold molar excess of CuSO4 for 30 min at room temperature. To remove excess copper, the protein was loaded onto a desalting column equilibrated with 20 mM Tris-HCl pH 8.0. 14 ul protein solution at 48 mg/ml was mixed with 14 ul reservoir buffer consisting of 1.9 M DL-malic acid pH 7.0