| Assay Method Information | |
| | Fluorescent Assay |
| Description: | The assays were performed by mixing either 0.2 μM E. coli OPPS, Y107A/F108A S. cerevisiae GGPPS, or Y96A/F97A human GGPPS with various concentrations of MANT-O-GPP and cold IPP as specified. For MANT-O-GPP Km measurements, 0.5−5 μM MANT-O-GPP was used with 50 μM cold IPP. For IPP Km and kcat determination, 3.5 μMMANT-O-GPP was utilized to saturate the enzyme with cold IPP from 2 to 50 μM. A standard curve was generated to correlate the fluorescence changes upon incubation with 0.2 μM enzyme, 0.5, 1, 2, 3.5, or 5 μM MANT-O-GPP, and 50 μM cold IPP for 30 min until the reaction reached completion. |
| Affinity data for this assay | |
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