EC Number   |
|---|
    3.5.3.1 | - |
| 3.5.3.1 | 12-16 mg/ml purified recombinant wild-type enzyme and mutant H141C complexed with substrate L-arginine, reaction intermediate Nomega-hydroxy-L-arginine and analogue Nomega-hydroxy-nor-L-arginine, hanging drop vapour diffusion method, equal columns of protein and precipitant solution containing 50 mM bicine, pH 8.5 at 22°C, 12-18% PEG 8000, 5 mM MnCl2, equilibration at 4°C over 1 ml precipitant solution as reservoir, pyramidal crystals after 4 weeks, complex preparation by soaking of crystals in 18% PEG 8000, 50 bicine, pH 8.5, 5 mM MnCl2, and 5 mM of Nomega-hydroxy-L-arginine and L-arginine for 6 days, complex formation with analogue Nomega-hydroxy-nor-L-arginine affords equilibration to pH 7.5, X-ray diffraction structure determination and analysis at 2.0-2.9 A resolution |
| 3.5.3.1 | 7 mg/ml purified fully active, untruncated enzyme complexed with 5 mM inhibitor S-(2-boronoethyl)-L-cysteine in 50 mM bicine, pH 8.5, 0.003 ml mixed with 0.1 ml MnCl2 and 0.003 ml precipitant solution containing 0.1 M Tris-HCl, pH 8.3-8.5, 3 M ammonium sulfate, equilibration over 1 ml reservoir solution containing 0.1 M Tris-HCl, pH 8.3-8.5, 3 M ammonium sulfate, and 20% v/v glycerol, hanging drop vapour diffusion method at 4°C, X-ray diffraction structure determination and analysis at 2.7 A resolution |
| 3.5.3.1 | crystallization by sitting-drop vapor diffusion method at 21°C. Crystal structures of the enzyme complexed with alpha,alpha-disubstituted boronic amino-acid inhibitors |
| 3.5.3.1 | D183A and D183N mutants of human arginase I complexed with 2(S)-amino-6-boronohexanoic acid are crystallized by the sitting drop vapor diffusion method at 21°C |
| 3.5.3.1 | homology modeling based on Bacillus caldovelox crystal structure. Residues D116, D120, D234, D236 and H91, H118, H133 contribute to catalysis and stability of binuclear metal center of arginase and have an important role in binding and catalytic activity in the active site |
| 3.5.3.1 | homology modelling and molecular dynamics study on the mechanism of metal dependency. When the active site metals are removed, loss of structural integrity is observed reflected by a larger equilibration root mean square deviation for the protein when the active site metal is removed and some loss of secondary structure. An inter-monomer salt-bridge between Glu295 and Arg404 is associated with the metal dependency |
| 3.5.3.1 | in complex with inhibitors 2(S)-amino-6-boronohexanoic acid or S-(2-boronoethyl)-L-cysteine |
| 3.5.3.1 | in complex with inhibitors N-hydroxy-L-arginine and nor-N-hydroxy-L-arginine, to 2.04 and 1.55 A resolution, respectively, and in complex with L-lysine, to 1.9 A resolution. Enzyme forms hydrogen bond interactions with inhibitor alpha-carboxylate and alpha-amino groups |
| 3.5.3.1 | in complex with inhibitors, vapor diffusion method, using 100 mM malonic acid, imidazol, boric system pH 5, and 25% or 30% (w/v) polyethylene glycol 1500 for aginase I or II, respectively |
