7W7R

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

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID30B
Synchrotron site	ESRF
Beamline	ID30B
Temperature [K]	100
Detector technology	PIXEL
Collection date	2016-12-04
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.911647
Spacegroup name	C 2 2 21
Unit cell lengths	113.670, 178.100, 177.900
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	84.360 - 3.460
R-factor	0.2721
Rwork	0.271
R-free	0.29910
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1j4n
RMSD bond length	0.003
RMSD bond angle	0.704
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]	95.818	95.818	3.650
High resolution limit [Å]	3.460	10.940	3.460
Rmerge		0.089	0.832
Rmeas	0.202	0.114	0.942
Rpim	0.095	0.068	0.432
Total number of observations	107214	3353	14473
Number of reflections	23598	814	3367
<I/σ(I)>	4.5	12.9	1.3
Completeness [%]	98.8	97.4	97.9
Redundancy	4.5	4.1	4.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		277.15	reservoir solution: 0.3M Lithium sulfate, 0.1M ADA (pH 6.5), 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 Dextran sulfate sodium salt Mr 5000. 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)