EC Number   |
|---|
    1.4.1.4 | - |
| 1.4.1.4 | chimeric protein consisting of domain I from NAD+-dependent GDH of Clostridium symbiosum, residues 1-200, domain II from NADP+-dependent GDH of Escherichia coli, residues 201-404 and the C-terminal helix again from Clostridium symbiosum, residues 405-448 which re-enters domain I. Domain II maintains its structural and functional integrity independent of the hinge and domain I. The enzyme is fully functional and retains the preference for NADP+ cofactor from the parent E. coli domain II |
| 1.4.1.4 | cofactor binding domain of glutamate dehydrogenase, sitting-drop vapor diffusion method. X-ray structure of the domain of wild-type enzyme and mutant enzyme R190A/E231A/K193A is solved at 1.43 A |
| 1.4.1.4 | complexed with NADP+ and 2-iminoglutarate. Among six subunits of hexameric GDH-binding NADP+, only four subunits bind 2-iminoglutarate in a closed form, while the other two are in an open form. In the closed form, 2-iminoglutarate is bound to the substrate-binding site with the 2-imino group stacked by the nicotinamide ring of the coenzyme, suggesting a prehydride transfer state |
| 1.4.1.4 | crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol (PEG) 400 as the precipitant |
| 1.4.1.4 | dialysis against ammonium sulfate solution |
| 1.4.1.4 | hanging-drop method of vapour diffusion using an ammonium sulfate and PEG mixture as the precipitant. The crystals belong to the monoclinic system and are in space group C2 with unit-cell dimensions a = 142.7, b = 202.0, c = 125.8 A with beta = 113.1 degrees with a hexamer in the asymmetric unit |
| 1.4.1.4 | in complex with NADPH and 2-oxoglutarate. The enzyme functions as a hexamer, and each monomer comprises a Rossmann-fold cofactor-binding domain and a substrate-binding domain. In the apo-form, GDH exists as an open state, and upon binding of the substrate and cofactor the protein undergoes a conformation change to a closed state |
| 1.4.1.4 | plate-like crystals of monoclinic space group C2 grown by vapour-diffusion using the sitting-drop method, X-ray crystallography to a resolution of 2.7 A |
| 1.4.1.4 | purified GDH in the absence of reactants, hanging drop vapour diffusion method, mixing of 10 mg/ml protein in 20 mM Tris-HCl, pH 7.6, with 15-25% PEG 3350, Tris/HEPES, pH 70-8.0, and 0.2 M NaCl, 18°C, 2 days, X-ray diffraction structure determination and analysis at 2.5 A resolution, modelling |
