5O0N

Deglycosylated Nogo Receptor with native disulfide structure 4

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-1
Synchrotron site	ESRF
Beamline	ID23-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-04-27
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	1.0
Spacegroup name	P 41
Unit cell lengths	90.630, 90.630, 45.620
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	90.630 - 2.500
R-factor	0.2201
Rwork	0.219
R-free	0.24790
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1ozn
RMSD bond length	0.004
RMSD bond angle	1.123
Data reduction software	iMOSFLM
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX ((1.11.1_2575: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	90.630	2.610
High resolution limit [Å]	2.500	2.500
Number of reflections	13084
<I/σ(I)>	13.2	2.4
Completeness [%]	100.0	100
Redundancy	7.4	7.6
CC(1/2)	0.998	0.704

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	291	Co-crystallization trial with mouse Nogo54, an extracellular construct derived from the NgR ligand Nogo-A (UNIPROT Q99P72, residues 1025-1078). Nogo54 was purified in 150 mM NaCl, 20 mM HEPES pH 7.0 at a concentration of 3.3 mg/mL and mixed in a 1:1 molar ratio with Endo-Hf-deglycosylated NgRa at a concentration of 10 mg/mL. Crystals were grown in a condition of 0.2 M ammonium sulfate, 0.1 M Bis-Tris pH 6.5, 25% (w/v) PEG3350.