EC Number   |
|---|
    2.7.7.18 | - |
| 2.7.7.18 | crystal structure of NaMN-bound form at 1.7 A, ATP-bound form at 2.0 A, apo-form at 2.0 A. The substrate-unbound and substrate-complexed structures are all in the fully open conformation. There is little conformational change upon binding each of the substrates. A conformational change is necessary to bring the two substrates closer together for initiating the catalysis. The authors suggest that such a conformational change likely occurs only after both substrates are simultaneously bound in the active site |
| 2.7.7.18 | crystallization of the apo-enzyme to space group: P1 with 44% sovent, and in complex with deamido-NAD+ with 42.4% solvent in space group I222, hanging-drop vapour-diffusion method at 20°C in 100 mM Tris, pH 7, 200 mM NaCl and 800 mM sodium citrate |
| 2.7.7.18 | Crystals of saNaMNAT were initially obtained at 22°C from polyethylene glycol 3000 and phosphatecitrate buffer. |
| 2.7.7.18 | crystals of the apo-enzyme belong to space group: P21 with 58.5% solvent and contain four molecules of NaMN AT in the asymmetric subunit, crystals in complex with deamido-NAD+ belong to space group: P212121 with 55.6% solvent and contain six molecules of NaMN AT in the asymmetric subunit, conditions: polyethylene glycol 3350 and 100 mM MgCl2 |
| 2.7.7.18 | homology modeling of structure, based on structures of isoform NMNAT1 and NMNAT3 and molecular docking of potential inhibitors |
| 2.7.7.18 | in complex with inhibitors 4-[2-(anthracen-9-ylmethylidene)hydrazino]-N-(3-chlorophenyl)-4-oxobutanamide, 4,4'-[cyclohexa-2,5-diene-1,4-diylidenebis[(E)methylylidene(E)diazene-2,1-diyl]]bis[N-(2-chlorophenyl)-4-oxobutanamide], and [(2E)-1-[4-[(2-chlorophenyl)amino]-4-oxobutanoyl]-2-(naphthalen-1-ylmethylidene)hydrazino]acetic acid. 4-[2-(Anthracen-9-ylmethylidene)hydrazino]-N-(3-chlorophenyl)-4-oxobutanamide binds at an enzyme monomer-monomer interface formed in the crystal of the complex, it sits at a central cleft between strands beta1 and beta4 of the beta sheet, which is the catalytic and substrate binding sites of the enzyme. The structures reveal a common binding site near residues Trp117, Try112, and Met109. This site overlaps but is distinct from the substrate-binding pocket |
| 2.7.7.18 | purified recombinant wild-type and W117A mutant enzymes, sitting drop vapour diffusion method at 20°C, microcrystal seeding using hanging drops, mixing of 10 mg/ml protein in 20 mM Tris, pH 7.8, 200 mM NaCl, and 5% v/v glycerol with well solution containing 1.2-1.5 M MgSO4, 0.1 M MES buffer, pH 6.0-6.5, microseeds are added to the drops 1 h later, macrocrystals growth within 3-5 days., X-ray diffraction structure determination and analysis at 2.4 A resolution, molecular replacement |
| 2.7.7.18 | sitting drop and hanging drop vapour diffusion method, mixing 0.001 ml of 4 mg/ml protein in 50 mM Tris, pH 7.0, 100 mM NaCl, 100 mM Arg, 100 mM Glu, 1% glycerol, 1 mM DTT, and containing 200 mM trehalose, with 0.001 ml of reservoir solution containing 2.1 M ammonium sulfate, 100 mM HEPES pH 7.0, 2% PEG 400 and 10 mM MgCl2, cryoprotection by 25% glycerol, X-ray diffraction structure determination and analysis at 2.3 A resolution, usage molecular replacement and of self-interaction chromatography for process optimization |
| 2.7.7.18 | structure of isoform NMNAT3 to 2 A resolution |
