EC Number   |
|---|
    3.2.1.26 | - |
| 3.2.1.26 | 2.6 A crystal structure of cell-wall invertase 1 in complex with a protein inhibitor, cell-wall inhibitor of beta-fructosidase, from tobacco. The structure recognizes a small amino acid motif in the inhibitor that directly targets the invertase active site. The activity of INV1 and its interaction with inhibitor are strictly pH-dependent with a maximum at about pH 4.5. At this pH, isothermal titration calorimetry reveals that the inhibitor tightly binds its target with nanomolar affinity. Cell wall inhibitor of beta-fructosidase competes with sucrose for the same binding site |
| 3.2.1.26 | apo form of beta-fructofuranosidase and its complex with fructose, both to 1.8 A resolution. Enzyme consists of the N-terminal five-blade beta-propeller domain that includes the catalytic site, as well as the beta-sandwich C-terminal domain. The active site is located at the bottom of the axial funnel created by the interblade loops and the loops between beta-strand 2-3 of each blade. The protein has a 35-residue elongation of the N-terminus containing a fiveturn alpha-helix, which distinguishes it from the other known members of the GH32 family |
| 3.2.1.26 | both native and SeMet-substituted InvB are concentrated to 5-10 mg/ml by ultrafiltration for crystallization. Crystals are grown at 16°C using the hanging drop vapor diffusion method |
| 3.2.1.26 | hanging drop vapour diffusion method using 22.5-28% PEG 8000 or 25% PEG 4000 as a precipitant in 0.1 M cacodylate pH 6.5 and 0.1 M zinc acetate |
| 3.2.1.26 | hanging drop vapour diffusion method, in complex with cell-wall inhibitor of beta-fructosidase, at pH 7.0 with sodium formate as precipitant, with PEG 4000, NaI at pH 7.5-9.5 or with PEG 4000 and Li2SO4 at pH 4.6 and 5.0, respectively |
| 3.2.1.26 | partial proteolysis and multiple sequence alignment indicate that both N and C termini are somewhat unstructured. Thus, we overexpressed and purified a truncated version of InvA (residues Lys9-Thr460) and successfully optimized its crystal co-crystallized with 200 mM fructose, the 2.11 A SeMet-substituted crystal of which was used for structure determination. In the sucrose-complexed structure (InvA-Suc), which has a space group of C2221, each asymmetric unit contains three molecules of InvA |
| 3.2.1.26 | purified recombinant His-tagged wild-type and selenomethionine-labeled enzyme, vapour diffusion method, 11 mg/ml wild-type enzyme in 15% PEG 1000, 150 mM Li2SO4, and 100 mM sodium citrate, pH 4.2, 3 days, 20°C, conditions for the selenomethionine-labeled enzyme: 8 mg/ml protein in 17% PEG 1000, 50 mM Li2SO4, 1% isopropanol, and 100 mM sodium citrate, pH 4.2, X-ray diffraction structure determination and analysis at 2.0-2.2 A resolution |
| 3.2.1.26 | purified recombinant inactivated mutant enzyme E190D in complex with substrate raffinose, hanging drop vapour diffusion method, 11 mg/ml protein in 15-17% PEG 1000, 150 mM Li2SO4, and 100 mM sodium citrate, pH 4.2, soaking of crystals in 5 mM substrate solution, flash freezing at -173°C, X-ray diffraction structure determination and analysis at a.87 A resolution, structure modelling, no successful crystal structure analysis with crystals of wild-type enzyme and mutant E190A built by the same method |
| 3.2.1.26 | sitting drop vapor diffusion method, using 3% (w/v) PEG 3350, 5% (w/v) 2-methyl-2,4-pentanediol, 0.6 M magnesium formate, 1 mM tris(2-carboxyethyl)phosphine, 0.33 M guanidinium chloride, and 0.1 Bis-Tris (pH 6.5) |
