Case Study

Part I – A lecture on enzyme inhibition

It was about 50 minutes into another long, boring two-hour lecture by Prof. P. The gray-haired biochemistry
professor was delivering another lecture on enzymes. The topic was on enzyme inhibition.
M., an energetic and outgoing student, was daydreaming about her upcoming weekend at home for a family
reunion. She was not really paying attention, but heard the professor going on and on about alcohol
dehydrogenase. At last, Prof. P. turned to the class and asked, “Any questions?”
M., hoping to side-track the professor, enthusiastically waved her hand. As soon as she received a nod from Prof.
P., she began to recount a story that her grandfather had told her.
“My grandfather said that when he was a teenager, alcohols were used as antifreeze in automobiles. A group
of kids was hanging out in a friend’s garage. One of them found two bottles of liquid that were marked
‘alcohol.’ He talked everyone else into trying the alcohol. All of the kids became severely ill, but about half
of the teens died from poisoning. Someone later found out that one of the bottles contained methanol and
the other a mixture of ethanol and methanol. The kids who lived drank the mixture. Grandpa figured since
I was a chemistry major, I’d know why half died and half didn’t, but I didn’t know. Do you?”
Although M. had planned to side-track the professor, her story played nicely into his lesson plan. After all, the
incident was related to the current class topic of enzymes and enzyme inhibition. The clever professor began,
“Let me remind all of you that the enzyme “X” catalyzes the first oxidation step in the case of both ethanol and
methanol. Methanol is a very common solvent that is quite toxic.
I recall an article from some years ago title “An Outbreak of Acute Methyl Alcohol Intoxication”, here is
part of the abstract:
“An outbreak of acute methyl alcohol intoxication occurred in Port Moresby, Papua New Guinea, in
March 1977. Twenty‐eight young men attended a drinking party and drank methyl alcohol. The amount
consumed by each individual ranged from an equivalent of 60–600 ml of pure methanol. Three had
prior ethanol ingestion. All 28 became ill 8–36 hours after drinking and were hospitalized. The most
commonly observed clinical syndromes were: acute metabolic acidosis, severe visual impairment and
acute pancreatitis. Four died within 72 hours after admission to the hospital. All had severe metabolic
acidosis and visual impairment and three pancreatitis. Of 24 who recovered, 16 showed no residual
complications, six had bi‐lateral visual impairment and two had difficulty in speech as well as visual
impairment….”

Questions
1. Write the reactions mentioned above, including the structure of the products for the oxidation of both
methanol and ethanol. Name those products as well as the name of the enzyme responsible
(mentioned as X in text). Refer to your textbook and notes as needed.
2. Discuss possible reasons for why some of the teenagers in the grandfather’s story died (as well as some
from the Port Moresby 1977 intoxication) and the others grandfather’s story did not.
Part II – The experiment
To illustrate the story told earlier, Prof. P. set up an experiment with his students studying a system with the same
type of enzyme inhibition as observed in alcohols. They collected the following data:

I II III
Rate (V0) of methanol oxidation by
Methanol Rate (V0) of methanol oxidation by enzyme X (mM/min)
concentration (mM) enzyme X (mM/min)
in presence of 5.0 μM of Ethanol
0.5 23.5 16.67
1.0 32.2 25.25
1.5 36.9 30.49
2.5 41.8 37.04
3.5 44.0 38.91

Questions (continued)
3. Using the data above, draw Lineweaver-Burk plots (two curves on the same graph)
a. first, graph 1/V0, the inverse of the reaction velocity (rate of oxidation reaction) from the data in column II
versus the inverse of the methanol concentration in column I.
b. then, on the same graph, plot the inverse of the reaction velocity (rate of oxidation reaction) from the data
in column III versus the inverse of the methanol concentration in column I.
4. Describe the conclusions that can be supported from the data. What type of enzyme inhibition is
evident from the plot?
5. Suggest ways that the conclusions might explain the case described in the grandfather’s story. What
can be considered the substrate of the enzyme “X” and what would be the inhibitor?