Biochemistry I - Activity of PFK +/- AMP: PFK-DL1

In the following dry lab you will measure the activity of PFK as a function of ATP concentration in the absense or presence of AMP. The amount of F6P (substrate) will be fixed at 100 uM.

  1. Before starting, review the reaction catalyzed by PFK in glycolysis. What are the substrates and what are the products?
  2. Vary [ATP] and record the initial velocity in the absence of AMP (left box) and in the presence of 25 uM AMP (right box). Plot the initial velocity versus [ATP].Use ATP levels from 0 to 700 uM, in steps of 50 uM for the first few points, then steps of 100, and then steps of 200, e.g. 0, 50, 100, 150, 200, 300, 400, 600 ...
1. Enter values for [ATP] in the left box (the amount of AMP is fixed at 25 uM).
[ATP] = uM [ AMP ] = 25 uM  
2. For the above values of [ATP] and [AMP], Calculate vi
vi = uM/min([AMP]=0) vi = uM/min ([AMP]=25uM)
(Each calculated vo value has a small "experimental error" added to it.)