Answer (b), (e), (g).

Enzymes do not change a reaction’s equilibrium constant and thus catalyze

the reaction in both directions, making (b) and (g) correct. Enzymes increase the rate of
a reaction by lowering the activation energy, hence (e) is correct.

An enzyme is discovered that catalyzes the chemical reaction

SAD HAPPY

A team of motivated researchers sets out to study the enzyme, which they call happyase. They find that
the kcat for happyase is 600 s-1. They carry out several experiments.

When [Et] = 20 nM and [SAD] = 40 μM, the reaction velocity, V0, is 9.6 μM s-1. Calculate Km for the
substrate SAD.

Solution: We know kcat, [Et], [S], and V0. We want to solve for Km. We substitute kcat[Et] for Vmax in the
Michaelis-Menten equation.
Once you have worked with this equation, you will recognize shortcuts to solve problems like this. For
example, one can calculate Vmax knowing that kcat[Et] = Vmax (600 s-1 × 0.020 μM = 12 μM s-1 in this
case). A simple rearrangement of MM by dividing both sides by Vmax gives

Thus, the ratio V0/Vmax = 9.6 μM s-1/12 μM s-1 = [S]/(Km + [S]). This simplifies the process of solving
for Km, giving 0.25[S] or 10 μM.

In a separate happyase experiment using [Et] = 10 nM, the reaction velocity, V0, is measured as 3 μM s-
1
. What is the [S] used in this experiment?

Solution: We see that the Vmax for this enzyme concentration is 6 μM s-1. Note that the V0 is exactly half
of the Vmax. Recall that Km is by definition equal to the [S] where . Thus, the [S] in this
problem must be the same as the Km, or 10 μM. If V0 were anything other than , it would be
simplest to use the expression V0/Vmax = [S]/(Km + [S]) to solve for [S].
The researchers working on happyase discover that the compound STRESS is a potent competitive
inhibitor of happyase. Addition of 1 nM STRESS increases the measured Km for SAD by a factor of
2. What are the values for α and α′ under these conditions?

Solution: Recall that the apparent Km, the Km measured in the presence of a competitive inhibitor, is
defined as αKm. Because Km for SAD increases by a factor of 2 in the presence of 1 nM STRESS, the value
of α must be 2. The value of α′ for a competitive inhibitor is 1, by definition

In an experiment, an enzyme converts a substrate (S) to a product. The Km of the substrate for the
enzyme is 5 μM. An added competitive reversible inhibitor is found to have an IC50 value of 50 nMfor
the enzyme when the substrate concentration is 25 μM. Use the Cheng-Prussoffequation to calculate
the Ki of the inhibitor in nMunits.

Use the Cheng-Prussoff equation to answer the question