5E23

Human transthyretin (TTR) complexed with (2,7-Dibromo-fluoren-9-ylideneaminooxy)-acetic acid

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	2014-01-30
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.870060
Spacegroup name	P 21 21 2
Unit cell lengths	43.222, 85.961, 63.348
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	42.981 - 1.410
R-factor	0.1727
Rwork	0.171
R-free	0.19730
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4pm1
RMSD bond length	0.007
RMSD bond angle	1.296
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	REFMAC (Rigid body)
Refinement software	PHENIX (1.9_1692)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.996	1.460
High resolution limit [Å]	1.410	1.410
Rmerge	0.088	1.272
Number of reflections	46322
<I/σ(I)>	17.26	1.75
Completeness [%]	99.9	100
Redundancy	9.09	8.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.5	293	Protein: 10mg/ml in .1 M NaCl, .050 M Na acetate, pH 5.5. Complex: 3 micro-L TTR + 3 micro-L 0.02% azide + 1 micro-L inhibitor (.001 M) Precipitant: 18% PEG4K, 9% PEG10K, 0.12 M imidazole malate pH 6.0, 0.04 M ammonium acetate, pH 4.5. Cryoprotectant: 10 % diethylene glycol + 5 % MPD + 15 % 2,3-butanediol + 5 % 1,4-dioxane, 12.5% MPEG 5K, 0.1 M mixed ( Na acetate, ADA, Bicine) 60% pH 4 / 40% pH 9)