8BIK
Crystal structure of human AMPK heterotrimer in complex with allosteric activator C455
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | DIAMOND BEAMLINE I02 |
Synchrotron site | Diamond |
Beamline | I02 |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2016-04-29 |
Detector | DECTRIS PILATUS 6M-F |
Wavelength(s) | 0.97949 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 75.313, 127.975, 139.215 |
Unit cell angles | 90.00, 92.91, 90.00 |
Refinement procedure
Resolution | 69.520 - 2.500 |
R-factor | 0.183 |
Rwork | 0.181 |
R-free | 0.21500 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | pdbid 4CFF |
RMSD bond length | 0.010 |
RMSD bond angle | 1.130 |
Data scaling software | Aimless (0.5.23) |
Phasing software | PHASER |
Refinement software | BUSTER (2.11.7) |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 75.220 | 75.220 | 2.560 |
High resolution limit [Å] | 2.500 | 11.180 | 2.500 |
Rmerge | 0.069 | 0.040 | 0.863 |
Rmeas | 0.082 | 0.050 | 1.027 |
Rpim | 0.045 | 0.029 | 0.552 |
Total number of observations | 300129 | 3295 | 22438 |
Number of reflections | 91016 | 1071 | 6689 |
<I/σ(I)> | 8.9 | 26 | 1.4 |
Completeness [%] | 99.9 | 99.7 | 99.9 |
Redundancy | 3.3 | 3.1 | 3.4 |
CC(1/2) | 0.988 | 0.938 | 0.546 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION | 7.2 | 293 | AMPK a2b1g1 (5 mg/ml: 38 uM) was combined with AMP (4-fold excess), staurosporine (1.2-fold excess) and activator 455 (3-fold excess): and crystallised at 20 C by mixing 2 ul of the protein solution with 1 ul of 8 % PEG3350, 0.3 M guanidine hydrochloride, 0.1 M PIPES buffer pH 7.2. |