EC Number   |
|---|
    1.14.14.154 | building up of homology models based on crystal coordinates of enzyme in complex with inhibitors 4-phenylimidazole or fluconazole, modeling of substrate 24-methylene-24,25-dihydrolanosterol into active site |
| 1.14.14.154 | crystals are grown using a hanging drop vapor diffusion technique. Enzyme complexes with posaconazole and VT-1161 are crystallized in the monoclinic C121 space group, and the structures are refined to 2.86 and 2.0 A. X-ray structures of Candida albicans CYP51 complexes with posaconazole and VT-1161, providing a molecular mechanism for the potencies of these drugs |
| 1.14.14.154 | CYP51 in complex with 4,4'-dihydroxybenzophenone, enzyme in 20 mM Tris-HCl, pH 7.5, 200 mM NaCl, and 0.5 mM EDTA, is mixed with 4,4'-dihydroxybenzophenone, which is dissolved in Me2SO at 100 mM stock concentration, to final concentrations of 0.2 mM for protein and ligand resulting in needle-like crystals, larger crystals are obtained by hanging drop vapor diffusion method from 1.2 M lithium sulfate, 0.1 M HEPES, pH 7.5, and 2% isopropyl alcohol, cryoprotection by 20% glycerol, X-ray diffraction structure determination and analysis at 1.95 A resolution |
| 1.14.14.154 | for crystallization purposes, an N-terminal truncated construct is used where the membrane anchor sequence is replaced with the 5-amino acid sequence fragment MAKKT-. Crystals are grown at 23°C using the hanging-drop vapor diffusion method |
| 1.14.14.154 | hanging drop vapor diffusion method, using 0.1 M Tris-HCl, pH 8.0, and 9% (w/v) PEG 6000 |
| 1.14.14.154 | in complex with fluconazole |
| 1.14.14.154 | purified recombinant C-terminally His-tagged enzyme, for crystallization purposes, the N-terminal transmembrane domain upstream of Pro32 is replaced with MAKKTSSKGKL- in the construct used for co-crystallization with fluconazole and (R)-4'-chloro-N-(1-(2,4-dichlorophenyl)-2-(1H-imid-azol-1-yl)ethyl)biphenyl-4-carboxamide, and with MAKKT-(5'-ATGGTCAAGAAAACG-3') in the complex with posaconazole, hanging drop vapour diffusion method, 25°C, from 0.260 mM cytochrome P450 solution in 20 mM potassium phosphate buffer, pH 7.2, containing 200 mM NaCl, 0.1 mM EDTA,10% glycerol, and 0.048mMn-tridecyl-beta-D-maltoside preincubated with 1.2fold molar excess of posaconazole, or (R)-4'-chloro-N-(1-(2,4-dichlorophenyl)-2-(1H-imid-azol-1-yl)ethyl)biphenyl-4-carboxamide, and 2-fold molar excess of fluconazole against an equal volume of well solution containing 0.2 M potassium formate, pH 7.2, or 0.2 M sodium formate, pH 7.4, in the case of (R)-4'-chloro-N-(1-(2,4-dichlorophenyl)-2-(1H-imid-azol-1-yl)ethyl)biphenyl-4-carboxamide, and 15% w/v PEG 3550, X-ray diffraction structure determination and analysis, modelling |
| 1.14.14.154 | purified recombinant enzyme, crystal structure determination and analysis, overview |
| 1.14.14.154 | recombinant mutant C37L/C442A in complex with alpha-ethyl-N-4-pyridinyl-benzeneacetamide, protein is mixed with a ligand dissolved in DMSO at a 100 mM concentration to obtain a final protein concentration of 0.2 mM and a final ligand concentration of 1 to 5 mM, 15-30% PEG 4000, 2-12% isopropanol, 0.1 M HEPES, pH 7.5, X-ray diffraction at 1.53 A resolution |
| 1.14.14.154 | X-ray crystal structures of hexahistidine-tagged Saccharomyces cerevisiae lanosterol 14alpha-demethylase in complex with its substrate lanosterol, the pseudosubstrate estriol and the triazole drugs itraconazole, posaconazole, fluconazole and voriconazole |
