Phosphofructokinase is a four‑subunit protein with four active sites. Phosphofructokinase catalyzes step 3 of glycolysis, converting fructose‑6‑phosphate to fructose‑1,6‑bisphosphate. Phosphoenolpyruvate (PEP) is the product of step 9 of glycolysis. The PEP concentration in the cell affects phosphofructokinase activity. Select the true statements about PEP regulation of phosphofructokinase. 1. PEP is a feedback inhibitor of phosphofructokinase.
2. The apparent affinity of phosphofructokinase for its substrate increases when PEP binds.
3. PEP is a positive effector of phosphofructokinase.
4. PEP inhibition of phosphofructokinase yields a sigmoidal velocity versus substrate curve.
5. PEP competes with fructose-6-phosphate for the active site of phosphofructokinase.
6. The binding of PEP to one phosphofructokinase subunit causes a conformation change that affects the ability of the substrate to bind to the other subunits.

1. PEP is a feedback inhibitor of phosphofructokinase.

4. PEP inhibition of phosphofructokinase yields a sigmoidal velocity versus substrate curve.

6. The binding of PEP to one phosphofructokinase subunit causes a conformation change that affects the ability of the substrate to bind to the other subunits.

Explanation:

Phosphofructokinase-1, PFK-1, is an allosteric enzymes composed of four protein subunits.

Allosteric enzymes are enzymes that function through non-covalent binding of allosteric modulators which may be activators or inhibitors. They produce a characteristic velocity versus substrate sigmoidal curve. PFK-1 has a separate binding site for its substrate, fructose-6-phosphate and it's allosteric modulators: ATP, ADP or phosphoenolpyruvate, PEP.

The enzyme can exist in two conformations, the T-state (tense) or the R-state (resting). Binding of substrate causes a conformational change from T-state to R-state, whereas binding of allosteric inhibitors returns it to the T-state.

PEP, the product of step 9 in glycolysis, is an allosteric inhibitor of PFK-1. When it binds to the the allosteric site, it leads to conformational changes in PFK-1 from the R-state to the T-state which reduces the enzymes ability to bind the substrate. These changes are responsible for the sigmoidal velocity/substrate curve in allosteric enzymes.

Therefore, the true statements from the options above are 1, 4, 6.

Options 2,3 and 5 are wrong because PEP is a negative effector of PFK-1, thus its binding reduces the affinity of PFK-1 for its substrate. Also, PFK-1 being an allosteric enzyme has separate binding sites for its substrate and its modulators. Thus, there is no competition for active site binding by substrate and modulators.