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. |
