Biochemistry I - HIV Integrase Inhibitors

You can add any concentration of substrate and inhibitor to the reaction tube. a) Obtain initial velocity (vi) versus S data with no inhibitor (green box), and in the presence of the inhibitor (yellow). You want to add enough inhibitor to show an effect (try 2 nM). For [S], use values from 0.5 to 20. b) Use the resulting values of vo to calculate 1/vo and 1/[S]; c) Plot the data (both without and with added inhibitor) on a velocity curve and a double reciprocal plot. d) Based on these plots, what type of inhibitor is it? Competitive or mixed-type? e) Calculate KI = [I]/(α-1). f) Calculate K'I = [I]/(α'-1), if appropriate. 1. Enter values for [S] and [ I ] in these boxes. Remember to keep the inhibitor concentration the same for all [S]: [S] = uM [ I ] = nM 2. For the above values of [S] and [I], Calculate vi vi = uM/min (NoInh) vi = uM/min (Each calculated vi value has a small "experimental error" added to it.) Hint: Ignore the data <10% and >90% Vmax when drawing a line. These values will have more relative error and they are not necessary to determine an accurate slope and intercept on the reciprocal plot.