EC Number   |
|---|
   4.1.99.13 | determination of a crystal structure of Drosophila melanogaster (6-4) PL (Dm64) bound to a 15-mer DNA duplex containing a central T(6-4)T lesion |
| 4.1.99.13 | hanging drop vapor diffusion at 4°C |
| 4.1.99.13 | purified recombinant His-tagged enzyme, hanging drop vapor diffusion method, mixing of 0.005 ml of protein in 12.5 mM Tris (hydroxymethyl) amino methane, 1.25 mM EDTA, 2.5% v/v glycerol, 75 mM sodium chloride, pH 7.8, with 0.005 ml of reservoir solution containing 2-6% w/v PEG400, 100 mM 2-(N-morpholino) ethanesulfonic acid buffer, pH 6.0, 25°C, 3-7 days, X-ray diffraction structure determination and analysis at 1.45-1.95 A resolution |
| 4.1.99.13 | purified recombinant PhrBI51W mutant, is crystallized by hanging drop vapor diffusion method, mixing of 5 mg/ml protein in 12.5 mM Tris, 1.25 mM EDTA, 2.5% w/v glycerol, and 75 mM sodium chloride, pH 7.8, with reservoir solution containing 5% PEG 400 and 100 mM 2-(N-morpholino) ethanesulfonic acid, pH 6.5, in a 1:1 ratio, X-ray diffraction structure determination and analysis at 2.15 A resolution, molecular replacement using the wild-type PhrBWT structure, PDB ID 4DJA, as the initial search model. Purified recombinant PhrBY424F mutant is crystallized by sitting drop vapor diffusion method, mixing 0.004 ml of 4-6 mg/ml protein in 12.5 mM Tris, 1.25 mM EDTA, 2.5% w/v glycerol, and 75 mM sodium chloride, pH 7.8, with an equal volume of reservoir solution containing 5% w/v PEG 400, 100 mM 2-(N-morpholino) ethanesulfonic acid, pH 6.0, and equilibration against 1 mLl of reservoir solution at 16°C, in darkness, 3-7 days, X-ray diffraction structure determination and analysis at 2.50 A resolution |
| 4.1.99.13 | T(6-4)C lesion containing DNA duplex in complex with the (6-4) photolyase, by the hanging-drop vapour diffusion method, at 18°C, to 2.95 A resolution. Lesion is flipped out of the opened DNA duplex into the active site of the enzyme |
| 4.1.99.13 | to study how the enzyme recognizes the T(6-4)C and T(Dew)C lesion analogues in the active site, DNA duplexes are crystallized together with the (6-4) photolyase |
| 4.1.99.13 | two crystal structures of the (6-4) photolyase bound to lesion containing DNA before and after repair, repair does not involve oxetane formation before light-induced electron transfer. The histidine 369, supposed to activate the acylimine, is in a position that does not allow efficient proton donation and hence activation of this substructure |
