BME211 Engineering physiology, Fall 2022

Midterm exam (total 40 pts)

Submit your exam answers to a box located in front of Bldg 110 Rm 801-12.
Submission deadline is 6:00 pm, 10/21/2022. Late exam will not be accepted.

1. Michaelis-Menten Equation with uncompetitive inhibition (18 pts)

Here are schematic diagrams different types of inhibition.

For instance, uncompetitive inhibition can be modelled as follows:

𝑘1
→ 𝑘2
[𝐸] + [𝑆] [𝐸𝑆] → [𝐸] + [𝑃]
←
𝑘−1

𝑘3
→
[𝐸𝑆] + [𝐼] [𝐸𝑆𝐼]
←
𝑘−3

As in homework 1, our goal here is to derive Michaelis-Menten equation for uncompetitive

inhibition.
(a) Write down the kinetic equation for [𝐸], [𝑆], [𝐼], [𝑃], [𝐸𝑆] and [𝐸𝑆𝐼] using the law of
mass action. (3 pts)
𝑑[𝐸] 𝑑[𝐸𝑆] 𝑑[𝐸𝑆𝐼]
(b) Show that the total enzyme concentration remains unchanged, i.e., + + =
𝑑𝑡 𝑑𝑡 𝑑𝑡
0. Hence, again, we may define an initial enzyme concentration unambiguously as [𝐸]0 =
[𝐸] + [𝐸𝑆] + [𝐸𝑆𝐼]. (3 pts)
We are once again interested in quasi-steady state approximation of 𝑑[𝑃]⁄𝑑𝑡 by assuming
[ES] and [ESI] complexes are in instantaneous equilibrium, or 𝑑[𝐸𝑆]/𝑑𝑡 = 𝑑[𝐸𝑆𝐼]/𝑑𝑡 = 0.
𝑑[𝐸𝑆]
(c) By setting = 0, show that
𝑑𝑡

𝐾𝑀 [𝐸𝑆]
[𝐸] =
[𝑆]
where 𝐾𝑀 = (𝑘−1 + 𝑘2 )⁄𝑘1 is the Michaelis constant. (Hint: use the condition
𝑑[𝐸𝑆𝐼]⁄𝑑𝑡 = 0) (3 pts)
BME211 Engineering physiology, Fall 2022

𝑑[𝐸𝑆𝐼]
(d) By setting = 0, show that
𝑑𝑡

[𝐸𝑆][𝐼]
[𝐸𝑆𝐼] = ,
𝐾𝑖
where 𝐾𝑖 = 𝑘−3 ⁄𝑘3 is the dissociation constant for the inhibitor [𝐼]. (3 pts)
(e) Plug in the equation found in (c) and (d) into the equation [𝐸]0 = [𝐸] + [𝐸𝑆] + [𝐸𝑆𝐼].
Show that
[𝑆][𝐸]0
[𝐸𝑆] =
𝐾𝑀 + [𝑆][𝐼]/𝐾𝑖 + [𝑆]
𝑑[𝑃]
Finally, by substituting the above expression into = 𝑘2 [𝐸𝑆], show that
𝑑𝑡

𝑑[𝑃] (𝑉𝑚𝑎𝑥 /𝛼) ⋅ [𝑆]

=𝑣= ,
𝑑𝑡 𝐾𝑀 /𝛼 + [𝑆]
[𝐼]
where 𝑉𝑚𝑎𝑥 = 𝑘2 [𝐸]0 and 𝛼 = (1 + 𝐾 ). We see that for uncompetitive inhibition, the effect
𝑖
[𝐼]
can be seen as decreases of Michaelis constant and maximum velocity by the factor (1 + 𝐾 ).
𝑖
(3 pts)
If we take the reciprocal of the Michaelis-Menten equation, we obtain a linear equation in
1⁄[𝑆].
𝑉𝑚𝑎𝑥 [𝑆] 1 𝐾𝑀 + [𝑆] 𝐾𝑀 1 1
𝑣= → = = +
𝐾𝑀 + [𝑆] 𝑣 𝑉𝑚𝑎𝑥 [𝑆] 𝑉𝑚𝑎𝑥 [𝑆] 𝑉𝑚𝑎𝑥
The plot out of this equation is called the Lineweaver-Burk plot.
(f) Similar to what you have done in homework 1, how do x-intercept, y-intercept, and slope
change in the uncompetitive inhibition form of the Michaelis Menten equation? (3 pts)
BME211 Engineering physiology, Fall 2022

2. van't Hoff equation (12 points)

Consider a vertical tube with a cross-sectional area of 1 cm2. The bottom of the tube is closed
with a semipermeable membrane, and 1 gram of sugar is placed in the tube. The membrane-
closed end of the tube is then put into an inexhaustible supply of pure water at T = 300 K. What
will be the height of the water in the tube at equilibrium? (The weight of a sugar molecule is
3×10−22 g, and the density of water is 1 g/cm3). Let’s solve this step-by-step.

(a) Compute the number (𝑛𝑠𝑢𝑔𝑎𝑟 ) of sugar molecules in 1g of sugar in moles. (3 pts) (hint: use
Avogadro’s number, 𝑁𝐴 = 6.02 × 1023 𝑚𝑜𝑙 −1)
(b) Find the molar concentration (or molarity) (𝑐𝑠𝑢𝑔𝑎𝑟 ) of 1g of sugar dissolved in water in the
tube with the cross-sectional area, 1 𝑐𝑚2 and the height, ℎ (in 𝑐𝑚 ). (3 pts) (Hints:
Molarity is defined as the amount of solute in moles dissolved in a liter of water. 1 𝑀 (1
molar) = 1 𝑚𝑜𝑙/𝐿, 1𝐿 = 1000 𝑐𝑚3 )
(c) Find the hydrostatic pressure, Π (in 𝑎𝑡𝑚 , atmospheric pressure) underneath the water
column with the height, ℎ (𝑐𝑚). (3 pts) (Hint, the pressure is the force per area. e.g., 1 Pa
= 1 𝑁/𝑚2 and 1 𝑎𝑡𝑚 (atmospheric pressure) ≈ 105 𝑃𝑎. A general formula for pressure
is Π = 𝜌𝑔𝐿, where 𝜌 is the density of fluid, 𝑔 is the gravitational acceleration (𝑔 = 9.8
𝑚 ⋅ 𝑠 −2 ), and ℎ is the height. First, convert all units to those in S.I. such as meter and kg,
obtain the pressure in 𝑃𝑎, and then convert the unit to 𝑎𝑡𝑚.)
(d) Finally, obtain the final height, ℎ of the water in the tube due to osmosis by using van’t
Hoff equation, Π = 𝑐𝑅𝑇, where Π is hydrostatic pressure, 𝑐 is molar concentration, the
universal gas constant R = 0.082 𝐿 ⋅ 𝑎𝑡𝑚 ⋅ 𝑚𝑜𝑙 −1 ⋅ 𝐾 −1, temperature T = 300 𝐾 (3 pts).
(Hint: ℎ is between 300 𝑐𝑚 and 400 𝑐𝑚).