Assignment. A terrible plague of H3N2 influenza has begun to decimate the earth's population. You convince your boss at your pharmaceutical company to let you work on developing a new neuraminidase inhibitor to combat this new strain. You make several hundred potential inhibitors and assay them with the neuraminidase isolated from this new strain. Frustratingly, none of the compounds has the required Ki of less than 2 µM to effectively inhibit this enzyme. Finally, in desperation attempt, you decide to make one last compound. You then assay this compound with the viral neuraminidase (which is used in the assay at 1 nM concentration).
After doing a complete study at various substrate and inhibitor concentrations, you arrive at the following data. The numbers inside the box are the reaction rates measured in nM/s. This table is an embedded MS excel file that can easily be copied into Microsoft Excel by cutting and pasting.
1. Using the data given, determine the Vmax, kcat, and Km for the substrate with no inhibitor present (Be sure to include units!). Attach a rate vs. [S] plot and a Lineweaver-Burk plot below. Describe how you determined these parameters. Show equations.
2. Perform a complete inhibitor study (rate vs. [S] plots at each concentration of inhibitor, your plots should each have 5 lines) using Excel and the data within the table. Attach the rate vs. [S] and Lineweaver-Burk plots below and label each curve with the inhibitor concentration for the data shown by that curve.
3. Answer the following question. What type of inhibitor is this compound.
4. Using a secondary plot like we described in class, determine the Ki for this compound. Attach the secondary plot below and describe how you determined Ki from this plot.
5. Did your compound save the world from this outbreak? true or false.