7W7S

High resolution structure of a fish aquaporin reveals a novel extracellular fold.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, EMBL c/o DESY BEAMLINE P13 (MX1)
Synchrotron site	PETRA III, EMBL c/o DESY
Beamline	P13 (MX1)
Temperature [K]	100
Detector technology	PIXEL
Collection date	2016-12-19
Detector	DECTRIS PILATUS3 S 6M
Wavelength(s)	0.8
Spacegroup name	P 4 21 2
Unit cell lengths	80.060, 80.060, 95.280
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	61.294 - 1.900
R-factor	0.1792
Rwork	0.178
R-free	0.20140
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1j4n
RMSD bond length	0.014
RMSD bond angle	1.171
Data reduction software	iMOSFLM
Data scaling software	SCALA (3.3.22)
Phasing software	PHASER
Refinement software	PHENIX (1.10.1_2155)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	95.280	95.280	2.000
High resolution limit [Å]	1.900	6.010	1.900
Rmerge		0.066	0.657
Rmeas	0.114	0.076	0.686
Rpim	0.034	0.033	0.195
Total number of observations	305910	9501	43587
Number of reflections	25108	924	3588
<I/σ(I)>	15.1	37.5	4
Completeness [%]	100.0	100	100
Redundancy	12.2	10.3	12.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		277.15	reservoir solution: 0.1M Tris-HCl (pH 7.8). 5% w/v gamma-PGA (Na+ form, LM), 30% v/v PEG 400. Prior to setting up the crystallization drops, 4 micro liter of the reservoir solution was mixed with 1 micro liter 30% w/v D-Sorbitol. Crystallization drops were set up by mixing the reservoir/additive mixture with protein at 1:1 or 2:1 ratio and the drops were left to equilibrate against 0.5 milliliter reservoir at room temperature and crystal grew in cold room (4 degree)