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) |