Biochemistry I - HIV Inhibitors

Enzyme Inhibition Kinetics: You can add any concentration of substrate and inhibitor to the reaction tube. The inhibitor concentration is the same for all three drugs (A, B, C). a) Obtain initial velocity (vi) versus S data with no inhibitor (green box), and in the presence of the inhibitors (yellow). Suggested values are:    Inhibitor - 2 nM.    [S] - 0.5, 1, 2, 10, 20 You only need 3-4 values to fit the straight line on the double reciprocal plot. c) Use the resulting values of vo to calculate 1/vo and 1/[S]; d) Plot the data (both without and with added inhibitor) on a single double reciprocal plot. e) Determine the slope of each plot, and calculate α, which is the ratio of the slopes. f) Calculate KI = [I]/(α-1). 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 (A) vi = uM/min (B) vi = uM/min (C) (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.