2W51

Human mesencephalic astrocyte-derived neurotrophic factor (MANF)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-1
Synchrotron site	ESRF
Beamline	ID23-1
Temperature [K]	93
Detector technology	CCD
Detector	ADSC CCD
Spacegroup name	P 61
Unit cell lengths	96.500, 96.500, 35.050
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	19.870 - 2.800
R-factor	0.281
Rwork	0.280
R-free	0.30500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	CDNF N-TERMINAL DOMAIN
RMSD bond length	0.007
RMSD bond angle	0.978
Data reduction software	XDS
Data scaling software	XDS
Phasing software	MOLREP
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.900
High resolution limit [Å]	2.800	2.800
Rmerge	0.040	0.540
Number of reflections	32978
<I/σ(I)>	20.1	2.6
Completeness [%]	97.0	94
Redundancy	7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	277	NEEDLE CRYSTALS OF MANF WERE TYPICALLY OBTAINED WITHIN 1 TO 2 WEEKS AT 4 C OVER A RESERVOIR SOLUTION OF 100 MM SODIUM CACODYLATE BUFFER, PH 6.5, 0.2 M MGAC2 AND 12-18 % PEG 8000.