EC Number   |
|---|
    1.14.18.1 | - |
| 1.14.18.1 | crystal structure analysis |
| 1.14.18.1 | hanging drop vapor diffusion method |
| 1.14.18.1 | hanging drop vapor diffusion method, using 10% (w/v) PEG 4000, 100 mM sodium acetate buffer pH 4.6 and 5 mM HoCl3 |
| 1.14.18.1 | hanging drop vapor diffusion method, using 18% (w/v) PEG 8000, 0.2 M zinc acetate, 0.1 M cacodylic acid pH 5.6-6.1, at 16°C |
| 1.14.18.1 | purified enzyme bound with L-tyrosine or L-dopa and zinc ions, hanging drop vapour diffusion method, mixing of 0.002 ml or 2 mg/ml protein in 20mM Tris-HCl, pH 7.5, and 500 mM NaCl, with 0.002 ml reservoir solution containing 18% PEG 8000 and 0.1 M cacodylic acid, pH 6.5, and equilibration against 0.6 ml of reservoir solution, 20°C, soaking of the crystals in inhibiting ZnCl2 before soaking them in the substrate solution in order to trap the substrates within the active site of TyrBm in the crystal, X-ray diffraction structure determination and analysis at 2.2-2.5 A resolution |
| 1.14.18.1 | purified enzyme in its resting met form with a Cu-Cu distance of 4.0 A, X-ray diffraction structure determination and analysis |
| 1.14.18.1 | purified recombinant detagged latent tyrosinase, X-ray diffraction structure determination and analysis. In contrast to the crystals obtained with the enzyme isolated from the natural source which contains two different chains in the asymmetric unit (one latent and one active protein) and does only form in the presence of sodium hexatungstotellurate(VI) (Na6[TeW6O24] x 22 H2O), the recombinant enzyme forms crystals containing exclusively the latent tyrosinase. The latent chain of AbPPO4 isolated from Agaricus bisporu, PDB ID 4OUA, chain B, is aligned with the heterologously produced protein, PDB ID 5M6B, chain B |
| 1.14.18.1 | purified recombinant wild-type enzyme and mutants A239T and F259A, hanging drop vapour diffusion technique, mixing 0f 0.001 ml of 5-10 mg/ml protein solution with 0.002 ml reservoir solution containing 50 mM Tris-HCl, pH 7.0, 19-21% PEG 3350, at 20°C, 10-15 days, method optimization, X-ray diffraction structure determination and analysis at 1.35, 1.55 and 1.70 A resolution, respectively. Soaking of crystals with a monophenolic (tyramine) and a diphenolic (dopamine) substrate in 50 mM Tris-HCl, pH 7.5, 200 mM NaCl, 20% PEG 3350, 20-25% PEG 1500, using SDS as an activator in order to perform in crystallo activity tests |
| 1.14.18.1 | sitting drop vapor diffusion method using 28% (w/v) PEG 1500, 100 mM bis-Tris buffer, pH 5.0, at 18°C, and hanging drop vapor diffusion method using 24% (w/v) PEG 2000, 10 mM NaCl, 10 mM CaCl2, and 3% 6-aminocaproic acid in 100 mM bis-Tris buffer, pH 6.5, at 18°C |
