EC Number   |
|---|
    1.4.3.22 | complexes of amine oxidase with the inhibitors benzylhydrazine and tranylcypromine are refined at 1.86 and 1.65 A resolution, respectively. Both inhibitors form covalent adducts with the 2,4,5-trihydroxyphenylalanine quinone cofactor. Tyrosine residue 296, proposed to act as a gate to the AGAO active site, is in its open conformation |
| 1.4.3.22 | in complex with aminoguanidine, hanging drop vapor diffusion method, using 0.1M bis-Tris propane (pH 7.5), 20% (w/v) PEG3350, and 0.2 M sodium sulfate at 20°C |
| 1.4.3.22 | purified recombinant wild-type hDAO, hanging drop vapour diffusion, 200 nl protein solution, containing 10 mg/ml protein in 100 mM HEPES, pH 7.2, and 150 mM KCl, and 200 nl crystallant solution, containing 0.1 M MES, pH 6.5, and 12% w/v PEG 20000, are equilibrated over 0.075 ml reservoir solution at room temperature, eight weeks, method optimization using manually dispensed hanging-drop crystallization experiments containing 0.002 ml each of protein and crystallant equilibrated over 0.5 ml reservoir solution, best diffracting crystals grow using 0.1 M MES pH 6.1 and 12% w/v PEG at room temperature over a period of two months, X-ray diffraction structure determination and analysis at 2.1 A resolution |
| 1.4.3.22 | sitting-drop vapor-diffusion method |
| 1.4.3.22 | the structure of the native enzyme is determined by X-ray crystallography to a resolution of 1.8 A. The homodimeric structure has the archetypal amine oxidase fold. Two active sites, one in each subunit, are characterized by the presence of a copper ion and a topaquinone residue formed. Substrate binding pocket and entry channel of hDAO are distinctly different from other amine oxidases in accord with the different substrate specificities. The structures of two inhibitor complexes of hDAO, berenil and pentamidine, are refined to resolutions of 2.1 and 2.2 A, respectively. They bind noncovalently in the active-site channel. The inhibitor binding suggests that an aspartic acid residue, conserved in all diamine oxidases but absent from other amine oxidases, is responsible for the diamine specificity by interacting with the second amino group of preferred diamine substrates |
