BCH 361 Midterm Study Questions

1. How do enzymes enhance the rate of chemical reactions?

2. What is the structural basis for enzyme specificity?
3. What would be the result of an enzyme having greater binding energy for the substrate than for
the transition state?
4. Why does the activation energy not appear in the G of a reaction?
5. Transition-state analogs, which can be used as enzyme inhibitors and to generate catalytic
antibodies are often difficult to synthesize. Suggest a reason.
6. Consider the following reaction:

After the reactants and products were mixed and allowed to reach equilibrium at 25C, the
concentration of each was measured:

Compound Equilibrium concentration (M)

glucose-1-phosphate 0.01
glucose-6-phosphate 0.19

a. Calculate Keq and G

b. If you had a solution of 0.1M glucose-6-phosphate and added to it
phosphoglucomutase, what will be the final concentrations of glucose-1-phosphate and
glucose-6-phosphate?
7. The isomerization of dihydroxyacetone phosphate (DHAP) to glyceraldehyde-3-phosphate (GAP)
has an equilibrium constant of 0.0475 under standard biochemical conditions. Calculate G for
the isomerization. Then, calculate G for this reaction when the initial concentrations of DHAP
and GAP are 2 x 10-4 and 3 x 10-6, respectively. What do these values tell you about the
importance of G compared with that of G in understanding the thermodynamics of
intracellular reactions.
8. Suppose you are studying a reaction that has a forward rate of 10-4 s-1 and a reverse rate of 10-6
s-1. What is Keq for this reaction? What is G? If you added an enzyme that increases the rate
of the reaction by a factor of 100, what will Keq and G be?
9. What is the biochemical advantage of having a KM approximately the same as the substrate
concentration that is normally available to the enzyme?
10. What is the defining characteristic of an enzyme catalyzing a sequential reaction? A double-
displacement reaction? (How can you tell these apart?)
11. The affinity between a protein and its ligand is frequently reported as a dissociation constant,
KD. Does KM measure the affinity between the enzyme and the substrate? Under what
circumstances might KM be approximately the same as KD?
12. The E. coli pyrophosphatase catalyzes the hydrolysis of pyrophosphate to two molecules of
inorganic phosphate. This enzyme has a molecular weight of 120 kDa and consists of six
identical subunits. For this enzyme, one unit of activity is defined as the amount of enzyme that
 hydrolyzed 10mol of pyrophosphate in 15 minutes at 37C under standard assay conditions.
The purified enzyme has a vmax of 2800 units per mg of enzyme.
a. Determine the number of moles of substrate that are hydrolyzed per second per mg of
enzyme when [S] is much greater than KM.
b. How many moles of active sites are there in 1 mg of enzyme? (Assume each subunit has
its own active site).
c. What is the turnover number for this enzyme? What does this value tell you about the
enzyme?
13. What is feedback inhibition? What is the importance of feedback inhibition?
14. Succinylcholine is used as a fast-acting, short-duration muscle relaxant. It is a competitive
inhibitor of acetylcholinesterase, the inhibition of which causes paralysis. However,
succinylcholine is hydrolyzed by blood-serum cholinesterase, which has a much broader
substrate specificity than acetylcholinesterase.
a. Before administration of succinylcholine, blood levels of serum cholinesterase are
determined. Why is this a good idea?
b. What would happen if a patient had a serum cholinesterase activity of 10 units/L rather
than the 80 normally found?
c. Some patients have a mutant form of serum cholinesterase with a KM of 10 mM rather
than the normal 1.4 mM. How will this mutation affect the patient?