EC Number   |
|---|
    2.7.1.36 | complex with farnesyl thiodiphosphate, 2.5 A resolution. Significant farnesyl thiodiphosphate hydrolysis occurs under crystallization conditions, this results in detection of farnesyl thiophosphate in the structure of the binary complex. Binding sites for these metabolites overlap, with the phosphate of farnesyl thiophosphate nearly superimposed on ATP's phosphate and farnesyl thiophosphate's polyisoprenoid chain overlapping ATPΒs adenosine moiety |
| 2.7.1.36 | crystals of mevalonate kinase-MgATP complex are grown at 4°C using the sitting drop method by mixing equal volumes of an enzyme solution containing 13 mg/ml protein, 1 mM ATP and 2 mM MgCl2 and a precipitant solution containing 100 mM HEPES buffer, pH 7.5 and 17.5% polyethylene glycol 5000 monomethylester, crystals appear after 3 days, crystal structure at 2.4 A resolution |
| 2.7.1.36 | free enzyme and in complex with mevalonate, 1.75 A and 1.9 A resolution, respectively. The mevalonate binds in a deep cavity lined by highly conserved residues. His25 is key for binding and for discrimination of (R)- over (S)-mevalonate, with the main chain amide interacting with the C3 hydroxyl group of (R)-mevalonate, and the side chain contributing, together with Val202 and Thr283, to the construction of a hydrophobic binding site for the C3 methyl substituent. The C5 hydroxyl, where phosphorylation occurs, points towards catalytic residues, Lys18 and Asp155 |
| 2.7.1.36 | free enzyme, 2.5 A resolution. Modeling of complex with farnesyl thiophosphate |
| 2.7.1.36 | hanging drop vapor diffusion method, the crystal structure is determined at 2.4 A |
| 2.7.1.36 | hanging drop vapor diffusion method, using 0.32 M MgCl2, 0.08 M bis-Tris pH 5.5, 16% (w/v) PEG 3350, at 18°C |
| 2.7.1.36 | in complex with diphosphomevalonate and Mg2+, diffraction to 2.5 A resolution. Diphosphomevalonate functions as a partial bisubstrate analogue and elicits a ternary-complex like form of the enzyme |
| 2.7.1.36 | purified enzyme in complex with 5-diphosphomevalonate and Mg2+ bound to the active cleft, sitting drop vapour diffusion method, 0.002 ml of 6,0 mg/ml protein in 50 mM HEPES, pH 7.5, 50 mM KCl, 1 mM DTT, 10 mM adenosine 5'-(beta,gamma-imino)triphosphate, 12 mM MgCl2, and 0.5 mM diphosphomevalonate is mixed with 0.002 ml of precipitatnt solution containing 40% v/v PEG 400, and 200 mM sodium formate, at room temperature, X-ray diffraction structure determination and anaylsis at 2.5 A resolution, single-wavelength anomalous diffraction method |
| 2.7.1.36 | purified enzyme in mevalonate-bound and 5-phosphomevalonate-bound form, hanging drop vapor diffusion method, mixing of 11 mg/ml protein in 20 mM Tris pH 7.7, 10 mM MgCl2, and 10% glycerol, with 100 mM Bis-Tris pH 5.5, 200 mM sodium potassium tartrate, and 15-25% PEG 8000 or 10000, microseeding. Seeded drops contain 0.001 ml of protein with 1 mM (R)-mevalonate, 5 mM AMPPNP, and 5 mM MgCl2, and 800 nl crystallization solution, and 200 nl of the seed stock. Diffraction-quality plate crystals grow in crystallization solutions that contain 100 mM Bis-Tris, pH 5.5, 200-300 mM sodium potassium tartrate, and 15-25% PEG 8000 or 10000, X-ray diffraction structure determination and analysis at 2.1-2.46 A resolution, molecular replacement using the structure of the apo-MmMK from Methanosarcina mazei (PDB ID 4HAC) as the search model |
| 2.7.1.36 | purified recombinant wild-type and selenomethionine-labeled enzyme as apoenzyme and in complex with (R)-mevalonate, hanging drop vapor diffusion method, 0.002 ml of 7.5 mg/ml protein in 10 mM Tris-HCl, pH 8.5, 20 mM NaCl, and 1 mM DTT, in presence of 3-25 mM adenosine 5'-(beta,gamma-imino)triphosphate, 6-50 mM (R)-mevalonate, 10 mM Mg2+, mixing with reservoir solution containing 1.15 M sodium citrate, pH 6.2, 18°C, X-ray diffraction structure determination and analysis at 1.75-1.9 A resolution, single-wavelength anomalous dispersion, molecular replacement fails |
