EC Number   |
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   2.7.11.31 | crystal structure of AMPK beta1 subunit-carbohydrate-binding module in complex with the cyclic sugar beta-cyclodextrin shows that the domain consists of a beta-hairpin loop extending from a beta-sandwich containing two anti-parallel beta-sheets. The sugar ring is held in position by the beta-hairpin loop, which protrudes the ring with Leu-146 at its centre. Within the sugarbinding pocket an extensive network of hydrophobic stacking interactions, mediated by Trp100 and Trp133, and carbohydrate-protein hydrogen bonds are formed with five of the seven glucose units. Although Leu-146 is prominent in the beta7-beta8 hairpin and interacts extensively with beta-cyclodextrin, it is not essential for glycogen binding |
| 2.7.11.31 | crystal structure of the inactive, apo-form of AMPK alpha2 subunit N-terminal kinase catalytic domain (KCD, residues 10-278 inclusive), shows that it adopts a canonical bilobal structure with the active site forming a cleft between the two lobes. The small N-terminal lobe (residues 1-97) is composed of a five-stranded beta-sheet (beta1-beta5), with an alpha-helix (termed the C-helix) positioned between strands beta3 and beta4 and lying to one side of the beta sheet. Glu-64 within the C-helix is important for aligning the phosphates of ATP in the correct orientation for catalysis. A Gly-X-Gly-X-X-Gly P-loop motif connecting strands beta1 and beta2 is evident in the structure. This interacts with the beta phosphate group of ATP when the active site is occupied. The larger C-terminal lobe is predominantly (63%) alpha-helical (alphaD-alphaI) and contains determinants and structural features that dictate protein substrate binding. The two lobes are connected via a short, flexible hinge region that allows rotation of the two lobes relative to each other |
| 2.7.11.31 | crystal structures for full-length Snf4 |
| 2.7.11.31 | crystal structures for full-length Snf4. In the subunit crystal structure ADP can be co-crystallized and occupies site 2, the unoccupied site present in mammalian gamma1 |
| 2.7.11.31 | crystals grown by mixing proteins with ammonium formate. Crystal structure of phosphorylated kinase domain, to 2.9 A resolution. Phosphorylated kinase domain displays a closed conformation |
| 2.7.11.31 | crystals grown by mixing proteins with ammonium sulphate. Crystal structure of an unphosphorylated fragment of the AMPK alpha-subunit that contains both the catalytic kinase domain and an autoinhibitory domain, to 2.8 A resolution. The unphosphorylated kinase domain of catalytic kinase domain/autoinhibitory domain fragment adopts an open inactive conformation. Kinase domain/autoinhibitory domain is jammed into its active site, and the critical catalytic residues make several intra- and intermolecular contacts |
| 2.7.11.31 | purified AMPK alpha2 subunit in apoform and in complex with compound C inhibitor, for the apoform: hanging-drop vapour diffusion method, mixing of 2.8 mg/ml protein in 20 mM Tris-HCl, pH 8.5, 0.3 M NaCl, 10% glycerol, 2 mM DTT, 5 mM MgCl2, and 5 mM AMPPNP, with reservoir solution consisting of 0.1 M Tris-HCl pH 8.9, 15% 2-propanol, 0.1 M ammonium sulfate, and 16% PEG 4000, for the complexed form: sitting-drop vapour-diffusion method, mixing of 2.8 mg/mlprotien in 20 mM Tris-HCl, pH 8.5, 0.3 M NaCl, 10% glycerol, 2 mM DTT, 5 mM MgCl2, and 0.5 mM compound C, with reservoir solution consisting of 0.1 M Bis-Tris, pH 6.5, 1.5 M ammonium sulfate, and 0.1 M NaCl, 20°C, X-ray diffraction structure determination and analysis at 2.08-3.0 A resolution, molecular replacement, modelling, overview |
| 2.7.11.31 | purified subunit beta1 carbohydrate-binding module CBD in complex with glucosyl-beta-cyclodextrin, sitting-drop vapour-diffusion method, mixing of 0.001 ml of 13 mg/ml protein in 20 mM HEPES, pH 7.0, and 6 mM gBCD, with 0.001 ml of reservoir solution containing 0.2 M lithium sulfate, 25% w/v PEG 8000, and 0.1 M sodium acetate, pH 4.5, X-ray diffraction structure determination and analysis at 1.72 A resolution, molecular replacement |
| 2.7.11.31 | purified subunit beta2 carbohydrate-binding module, sitting-drop vapour-diffusion method, mixing of 0.001 ml of 13 mg/ml protein in 20 mM HEPES, pH 7.0, with or without 6 mM glucosyl-beta-cyclodextrin, with 0.001 ml of reservoir solution, which contains for the unliganded enzyme 0.17 M ammonium sulfate, 15% v/v glycerol and 25.5% w/v PEG 4000, or contains 0.2 M lithium chloride, 20% w/v PEG 6000 and 0.1 M sodium HEPES, pH 7.0 for the complex with gBCD, X-ray diffraction structure determination and analysis at 1.6-2.0 A reolution, molecular replacement |
