EC Number   |
|---|
    3.5.4.1 | - |
| 3.5.4.1 | 10 mg/ml purified recombinant His-tagged enzyme, vapour phase equilibration against 11-14% PEG 8000, 0.1 M HEPES, pH 7.3-7.7, 0.2 M MgCl2, in hanging drop geometry, flash-cooling in a buffer containing 30% v/v glycerol for cryoprotection, also crystallization of a seleno-methionine derivatized enzyme, X-ray diffraction structure determination and analysis at 1.5 A resolution |
| 3.5.4.1 | crystallization strategy named microseed matrix screening, differential chelation pattern of cations by acidic surfaces of proteins within crystal lattice as a critical parameter of crystal nucleation and growth |
| 3.5.4.1 | molecular dynamics simulation of free enzyme and in complex with cytosine, uracil and reaction intermediates |
| 3.5.4.1 | PDB ID 1UAQ, structure analysis and comparison to the quantum mechanical/molecular mechanical molecular dynamics simulation model, overview |
| 3.5.4.1 | purified recombinant wild-type and selenomethionine-labeled enzymes in 10% 2-propanol, 20% PEG 4000, 0.1 M Na HEPES, pH 7.5, at 4°C and at 22°C by hanging drop vapour diffusion method, 0.002 ml of both reservoir and protein solution are mixed in presence of 2-hydroxypyrimidine, 3-5 days to 1-2 weeks at 22°C, micro-seeding, X-ray diffraction structure determination and analysis at beyond 1.5 A resolution |
| 3.5.4.1 | recombinant A23L/D92E/V108I/I140L mutant bound to the transition state analogue, hanging drop vapour diffusion method, 2 days, crystals are soaked for 20 min in a mother liquor solution containing 2-hydroxypyrimidine concentrated 1.2:1 relative to protein, after soaking, the crystals are immediately transferred briefly to a cryo-buffer containing the 2-hydroxypyrimidine mother liquor plus 25% DMSO, X-ray diffraction structure determination and analysis at 2.3 A resolution, modelling and molecular replacement |
| 3.5.4.1 | the crystal structure of cytosine deaminase combined with the substrate uracil, PDB ID code: 1P6O, optimization in the water solvent at the ONIOM molecular dynamics study, overview |
| 3.5.4.1 | the structure of Zn-CDA is determined to a resolution of 1.7 A with phosphonocytosine a potent mimic of the putative tetrahedral intermediate bound in the active site |
| 3.5.4.1 | the wild type enzyme in complex with the mechanism-based inhibitors 4-(R)-hydroxyl-3,4-dihydropyrimidine or 5-fluoro-4-(S)-hydroxyl-3,4-dihydropyrimidine and the mutant enzyme V152A/F316C/D317G are crystallized by the sitting drop vapor diffusion method, using 10-15% (w/v) PEG 6K, 200 mM MgCl2 and 100 mM HEPES (pH 7-8) |
