EC Number   |
|---|
   2.1.1.255 | purified recombinant His6-tagged apo-GPPMT containing one molecule in the asymmetric unit, or in ternary complex with geranyl diphosphate and sinefungin, hanging drop method, mixing of protein solution containing 5 mg/ml in 50 mM sodium phosphate, pH 7.5, 300 mM NaCl and 10% glycerol, with S-adenosyl-L-methionine at a protein:S-adenosyl-L-methionine molar ratio of 1:7, 4°C, and further with reservoir solution containing 50 mM sodium cacodylate, pH 6.5, 1.4 M lithium sulfate, 30 mM MgCl2, and 1 mM spermine for GPPMT-GPP-SFG form I, or 50 mM sodium cacodylate, pH 6.7, 1.3 M lithium sulfate, 30 mM MgCl2, and 1 mM spermine for GPPMT-GPP-SFG form II, method screening, X-ray diffraction structure determination and analysis at 1.7-3.5 A resolution, molecular replacement method |
| 2.1.1.255 | purified recombinant His6-tagged wild-type enzyme in complex with S-adenosyl-L-methionine and monoterpenoid ligands geranyl-S-thiolodiphosphate, a substrate analogue, or geranyl diphosphate, sitting drop vapor diffusion method, 4°C, mixing of 0.001 ml protein solution conatining 6 mg/mL enzyme, 50 mM PIPES, pH 6.7, 20% glycerol, 5 mM BME, 15 mM MgCl2, 100 mM NaCl, 10 mM S-adenosyl-L-methionine, and 1 mM monoterpenoid ligand with 0.001 ml of precipitant solution containing 100 mM HEPES, pH 7.5, 25% polyethylene glycol 3350, and 200 mM (NH4)2SO4 for cocrystallization with geranyl-S-thiolodiphosphate, 100 mM Bis-Tris, pH 6.5, 25% polyethylene glycol 3350, and 200 mM (NH4)2SO4 for cocrystallization with geranyl diphosphate and equilibrated against a 0.1 ml reservoir of precipitant solution at 21°C, 1 day, X-ray diffraction structure determination and analysis at 2.05 A resolution |
| 2.1.1.255 | sitting-drop vapour-diffusion method |
