EC Number   |
|---|
    3.2.1.24 | 30 mg/ml purified recombinant Golgi alpha-mannosidase IA in 20 mM MES, pH 6.5, 150 mM NaCl, 5 mM CaCl2, and 0.75 M NDSB-201, crystallization of free enzyme by microbatch method using a precipitation solution containing 15-20% PEG 4000 at pH 4.5-6.5, cocrystallization of 1-deoxymannojirimycin bound to the enzyme by hanging drop vapour diffusion method at 37°C, 0.001 ml of a solution containing 200 mM 1-deoxymannojirimycin is mixed with an equal volume of crystallization solution containing 100 mM MES and 100 mM Tris-HCl, pH 6.0, and 25-35% PEG 4000, 2 days at 18°C, X-ray diffraction structure determination and analysis at 1.5 A resolution, structure modelling |
| 3.2.1.24 | crystal structure at 2.7 A resolution |
| 3.2.1.24 | crystal structure at 2.8 A resolution |
| 3.2.1.24 | hanging drop vapour diffusion method, crystallization of wild-type enzyme in complex with 5-fluoro-beta-L-gulosyl fluoride, mutant enzyme D341N in complex with 2-deoxy-2-fluoro-alpgha-D-mannosyl fluoride and mutant enzyme D341N in complex with 5-fluoro-beta-L-gulosyl fluoride |
| 3.2.1.24 | in complex with inhibitors mannoimidazole, glucoimidazole, N-octyl-6-epi-valienamine, gluco-hydroxyiminolactam, and [[(3S,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)piperidin-2-ylidene]amino] N-(4-chlorophenyl)carbamate |
| 3.2.1.24 | purified recombinant TM1851, sitting drop vapour diffusion method, optimal conditions: 0.001 ml of protein solution containing 5.3 mg/ml protein in 5 mM sodium phosphate and 150 mM NaCl, pH 6.8, mixed with an equal volume of reservoir solution containing 4% w/v PEG 6000, 50 mM sodium phosphate, pH 6.0, and 0.5 M NaCl, 1 day at 25°C, X-ray diffraction structure determination and preliminary analysis at 2.9 A resolution |
| 3.2.1.24 | purified recombinent enzyme free or in complex with the inhibitor swainsonine, sitting drop vapour diffusion method, mixing of 12 mg/ml protein in 100 mM Tris, pH 8.5, 1.5 M (NH4)2SO4 and 12% v/v glycerol, with reservoir solution, containing 3% v/v glycerol, 54% v/v Tacsimate, pH 7.0, and 2% v/v PEG 6000, also acting as the cryo-protectant, crystals of the swainsonine complex form are obtained by soaking SpGH38 crystals for ,16 h in mother liquor supplemented with 2 mM swainsonine, X-ray diffraction structure determination and analysis at 1.9 A and 2.6 A resolution, respectively |
| 3.2.1.24 | structual 3D analysis |
| 3.2.1.24 | vapor diffusion and micro-batch crystallization techniques, crystal structure in absence and presence of the anti-cancer agent swainsonine and the inhibitor deoxymannojirimycin |
| 3.2.1.24 | X-ray crystal structures obtained in apo-, inhibitor-bound, and substrate-bound forms provide both mechanistic and molecular insight into how the proteins, which adopts an (alpha/alpha)6fold, recognizes and hydrolyzes the alpha1,6-mannosidic bond by an inverting, metal-independent catalytic mechanism |
