EC Number   |
|---|
   2.7.1.190 | apo form and in complex with a bound antibiotic, tobramycin and kanamycin A, to 2.05 A, 1.8 A and 2.15 A resolution, respectively. Substrate binding induces conformational changes, involving rotational shifts of two distinct segments of the enzyme. The helical subdomain is important in substrate specificity |
| 2.7.1.190 | crystal structures of isoform APH(2'')-IVa, wild-type and mutant F95M, in complex with either adenosine or guanosine |
| 2.7.1.190 | enzyme APH(2'')-Ia crystals, grown with guanosine-beta,gamma-imidotriphosphate (GMPPNP) and a saturating concentration of magnesium, are soaked with ribostamycin, hanging drop vapour diffusion method, mixing of 0.001 ml ofp protein solution with 0.001 ml of reservoir solution containing 100 mM HEPES, pH 7.5, 120 mM MgCl2, 10% PEG 3350, and 10% glycerol, preincubation of the enzyme with 3 mM GMPPNP and 6 mM MgCl2, X-ray diffraction structure determination and analysis at 2.20-2.60 A resolution, model building |
| 2.7.1.190 | purified enzyme in complex with substrate G418 and sisomicin, mixing 0.001 ml of 6 mg/ml protein and 2.5 mM aminoglycoside in 50 mM HEPES, pH 7.5, and 10 mM MgCl2 with 0.001 ml of reservoir solution containing 12% PEG 3350 w/v, and 50-75 mM ammonium citrate, pH 7.4-7.9, 18°C, X-ray diffraction structure determination and analysis at 3.05 A and 2.35 A resolution, respectively, molecular replacement using chain A of the homologous APH(2'')-IVa-ADP complex (PDB ID 4N57) after removal of the ligand as search model, modeling |
| 2.7.1.190 | purified recombinant wild-type and mutant enzymes in complex with cofactor GTP, GDP, and especially GMPPNP, and substrates gentamycin C1, rbiostamycin, kanamycin A, neomycin B in diffenrent combinations, X-ray diffraction structure determination and analysis at 2.15-2.50 A resolution |
| 2.7.1.190 | ternary complex of APH(2'')-IIIa with GDP and kanamycin, the ternary complex s prepared by adding a tenfold molar excess of Mg2-GTP and kanamycin to the apo APH(2'')-IIIa F108L enzyme, followed by incubation of the complex at 4x02C for 2 h, the complex is crystallized from 30% PEG 4000, and 0.1 M Tris-HCl pH 8.5, X-ray diffraction structure determination at 1.34 A resolution, molecular replacement using the binary Mg2-GDP-APH(2'')-IIIa complex as the starting model (PDB ID 3tdw) |
| 2.7.1.190 | to 2.2 A resolution. Access to the ATP-binding template is blocked by a bulky tyrosine residue. Substitution of this tyrosine by a smaller amino acid opens access to the ATP template |
