EC Number   |
|---|
    1.7.2.4 | 1.6 A resolution |
| 1.7.2.4 | 2.4 A resolution |
| 1.7.2.4 | 2.4 A resolution, preliminary study |
| 1.7.2.4 | building of two models of the active site reveals two distinct mechanisms. In the first model, N2O binds to the fully reduced tetranuclear Cu4S core in a bent my-(1,3)-O,N bridging fashion between the CuI and CuIV centres and subsequently extrudes N2 while generating the corresponding bridged my-oxo species. In the second model, substrate N2O binds loosely to one of the coppers of the tetranuclear Cu4S core in a terminal fashion, i.e., using only the oxygen atom. Loss of N2 generates the same my-oxo copper core. The free energies of activation predicted for these two alternative pathways are close to one another and do not provide decisive support for one over the other |
| 1.7.2.4 | hanging drop vapour diffusion method |
| 1.7.2.4 | modeling of structure. The residues contributing to the semiocclusion of the T1 copper site are Trp355, Met389, and Met297. There is a negatively charged residue in the neighborhood of the T1 site, Glu296 |
| 1.7.2.4 | purified enzyme, sitting drop vapor diffusion method, for His6-tagged Ca2+-bound apo-N2OR enzyme: mixing of 0.0003 ml of 15mg/ml protein solution with 0.0003 ml of well solution containing PEG 3350, 0.2 M ammonium sulfate, and 0.1 M Bis-Tris-HCl, pH 5.5, for Strep-tagged Ca2+-free or -bound apo-N2OR: mixing of 0.0003 ml of 15mg/ml protein solution with 0.0003 ml of well solution containing 25% w/v PEG 3350, 0.2 M lithium sulfate monohydrate, and 0.1 M Tris-HCl, pH 8.5, 7 days, 25°C, X-ray diffraction structure determination and analysis, molecular replacement and modelling using structure PDB ID 3SBQ as template. No crystals are obtained for Ca2+-free His6-tagged apo-N2OR |
