6ZFQ

Structure of the catalytic domain of human endo-alpha-mannosidase MANEA in complex with bis-tris

This is a non-PDB format compatible entry.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I24
Synchrotron site	Diamond
Beamline	I24
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-07-09
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.97718
Spacegroup name	P 1 21 1
Unit cell lengths	42.812, 106.994, 43.884
Unit cell angles	90.00, 110.31, 90.00

Refinement procedure

Resolution	38.410 - 1.200
Rwork	0.141
R-free	0.18120
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6zfn
RMSD bond length	0.013
RMSD bond angle	1.820
Data reduction software	DIALS (1.10.2-g9c23cb36c-release)
Data scaling software	Aimless (0.7.1)
Phasing software	PHASER (2.8.2)
Refinement software	REFMAC (5.8.0258)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	38.410	1.220
High resolution limit [Å]	1.200	1.200
Rmerge	0.049	0.695
Rmeas	0.057	0.854
Rpim	0.028	0.485
Number of reflections	114498	5426
<I/σ(I)>	10.6	1.5
Completeness [%]	99.4	94.7
Redundancy	3.8	2.8
CC(1/2)	0.999	0.804

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.5	292	100 mM bis-tris pH 5.5, 25% w/v PEG 3350. Protein at 10 mg/ml in 25 mM HEPES pH 7.0, 200 mM NaCl buffer with 2.23 mM GlcIFG and 2.23 mM alpha-1,2-mannobiose (10 x molar ratio). 300 nl droplet (150 nl protein solution, 150 nl reservoir solution). The ligand additives are probably not required for crystallization.