6WBS

Human CFTR first nucleotide binding domain with dF508/V510D

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2015-12-18
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.9786
Spacegroup name	P 21 21 21
Unit cell lengths	62.396, 81.660, 100.404
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	42.767 - 1.857
R-factor	0.1718
Rwork	0.170
R-free	0.21270
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2bbs
RMSD bond length	0.010
RMSD bond angle	1.184
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	PHENIX (1.13_2998)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.930
High resolution limit [Å]	1.857	4.010	1.860
Rmerge	0.124	0.050	0.916
Rmeas	0.127	0.055
Rpim	0.055	0.023	0.410
Number of reflections	43865	4653	4284
<I/σ(I)>	7
Completeness [%]	100.0	99.9	100
Redundancy	6	5.6	5.7
CC(1/2)		0.997	0.741

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	277	CFTR NBD1 V510D in buffer A was crystallized by mixing equal amounts of protein at 6 mg/mL with 0.1 M Tris pH 7.6, 28% (w/v) polyethylene glycol 10,000. Crystallization was induced by streak-seeding using crystals grown in 0.1 M HEPES, pH 7.5, 25% (w/v) polyethylene glycol 550 monomethylether, and plates incubated at 4C. Crystals grew over several days and were frozen by quick dip in reservoir solution supplemented with 25% (w/v) ethylene glycol