EC Number   |
|---|
    2.4.1.133 | enzyme in the presence of manganese and UDP, hanging drop vapor diffusion method, using 100 mM Tris-HCl (pH 8.0), 1 M NaCl, 15% (v/v) MPD, and 5% (w/v) polyethylene glycol 6000 |
| 2.4.1.133 | hanging drop vapor diffusion method, using 100 mM imidazole (pH 6.5) and 750 mM sodium acetate or 100 mM HEPES buffer (pH 8.0), 100mM serinol, 1.0 M NaCl, 5% (w/v) PEG6000, and 10% 2,4-methanepentadiol |
| 2.4.1.133 | purified recombinant detagged enzyme in complex xylobiose, hanging drop vapor diffusion method, mixing of protein solution containing 10 mg/ml protein, 10 mM MnCl2, 10 mM UDP, and 10 mM xylobiose, with a reservoir solution containing 100mM HEPES, pH 8.0, 100mM serinol, 1.0 M NaCl, 5% PEG 6000, and 10% 2-methyl-2,4-pentanediol, crystals of enzyme mutants D318N and D211N protein in complex with UDP-Gal, by hanging drop vapor diffusion method, using a protein solution containing 10 mg/ml protein, 10 mM MnCl2, and 10 mM UDP-Gal, with the precipitating solution containing 100 mM HEPES, pH 8.0, 1.0 M NaCl, 5% PEG 6000, and 10% 2-methyl-2,4-pentanediol, X-ray diffraction structure determination and analysis |
| 2.4.1.133 | purified recombinant detagged wild-type and truncated enzymes, hanging drop vapor diffusion method, mixing of 15 mg/ml protein solution with reservoir solution containing 100 mM imidazole, pH 6.5, 5 mM MnCl2, and 750 mM sodium acetate, 18°C, X-ray diffraction structure determination and analysis, molecular replacement using the Drosophila melanogaster beta4GalT7 crystal structure, PDB ID 3LW6, as a search model. For crystal complexes, tetragonal crystals are soaked in the same pH 6.5 reservoir solution containing 15% 2-methyl-2,4-pentanediol at 18 °C for 20 h followed by a brief soak in 100 mM Tris, pH 8.0, 750 mM sodium acetate, 15% 2-methyl-2,4-pentanediol, 5 mM MnCl2, and 5 mM UDP-Gal at room temperature. The monoclinic crystals are obtained within 3 days using 100 mM Tris, pH 8.5, and 8% PEG 8000 as a reservoir solution by mixing it with 20 mg/ml beta4GalT7DELTA81 in 10 mM MES (pH 6.5), 150 mM NaCl, 5 mM MnCl2, and 5 mM UDP-galactose at 18 °C |
