|Biochemistry I - Regulation of PFK by F26P (PFK-DL2)
(Each calculated vo value has a small "experimental error" added to it.)
In the following dry lab you will measure the activity of PFK as a function of F-6-P concentration in the absense or presence of various amounts of F26P. The concentration of ATP and AMP (Activator) are set to give the optimal activity for the enzyme - you will adjust only the F-6-P and F26P levels.
- Before starting, review the reaction catalyzed by PFK in glycolysis. What are the substrates and what are the products?
- How does the structure of the regulatory compound, F26P, differ from the substrates and products for PFK?
- Vary [F6P] and record the initial velocity of PFK in the absence of F26P(left box, yellow) and in the presence of 25 uM F26P (right box, orange).
- How does F26P affect the reaction velocity?
- Where do you think F26P binds to the enzyme, in the active site, or elsewhere?
- Use the effect on the velocity (F6P saturating) to measure the Kd for F26P binding.