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Cellular Respiration Virtual Lab Notes

1) The document describes a virtual lab experiment where a student named Jared was poisoned. Measurements were taken of pyruvate, NADH, and intermembrane hydrogen ion concentrations in Jared's muscle cells and compared to healthy muscle cells. 2) The data supported the hypothesis that Jared was poisoned with cyanide, which halts the electron transport chain, preventing the formation of a proton gradient and ATP production. Only cyanide caused the observed changes in metabolite concentrations. 3) An antidote was given that converted the cyanide to thiocyanate, allowing the electron transport chain to resume function and the production of ATP through the proton gradient.

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4K views4 pages

Cellular Respiration Virtual Lab Notes

1) The document describes a virtual lab experiment where a student named Jared was poisoned. Measurements were taken of pyruvate, NADH, and intermembrane hydrogen ion concentrations in Jared's muscle cells and compared to healthy muscle cells. 2) The data supported the hypothesis that Jared was poisoned with cyanide, which halts the electron transport chain, preventing the formation of a proton gradient and ATP production. Only cyanide caused the observed changes in metabolite concentrations. 3) An antidote was given that converted the cyanide to thiocyanate, allowing the electron transport chain to resume function and the production of ATP through the proton gradient.

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3 Cellular Respiration Virtual Lab 17 September 2020

Jason Bao
Cellular Respiration Virtual Lab

Control Data

Healthy Muscle Cells

Pyruvate 0.12 mM

NADH 0.30 mM

Intermembrane H+ 0.32 mM
Jared’s Cell Data

Pyruvate concentration stays constant.

NADH concentration increases.

Intermembrane H+ Concentration decreases.

Healthy Muscle Cells Jared’s Muscle Cells

Pyruvate 0.12 mM 0.12 mM

NADH 0.30 mM 0.50 mM

Intermembrane H+ 0.32 mM 0.05 mM


Hypothesis:
Choose the poison you think was given to Jared: Cyanide
Explain how the data supports your hypothesis: The data supports the hypothesis that Jared was
poisoned with cyanide as Jared’s muscle cells severely lack H+ in the intermembrane space,
protons that the Electron Transport Chain provide when NADH and FADH2 drop off their
electrons. However, since the data indicates a high concentration of NADH and a low
concentration of intermembrane H+, the ETC must have halted, which the poison cyanide does by
attaching to the last enzyme of the ETC.

Experiments: Measuring changes in concentration of pyruvate, NADH, and intermembrane H+


after each poison injected into muscle cells
2-deoxyglucose – all decrease
Arsenic – pyruvate increase, others decrease
Cyanide – pyruvate no change, NADH increase, H+ decrease
Oligomycin – pyruvate and NADH no change, H+ increase

Review:
Final Hypothesis – Cyanide was used to poison to Jared’s cells.
Antidote – antidote changes cyanide to molecule called thiocyanate, which the body can remove
through urine.

Case Summary:
Background
Explain how cyanide caused Jared’s symptoms (e.g., muscle weakness).
The cyanide attaches itself to the last enzyme of the ETC, which halts the processes of the chain.
As such, the ETC is unable to pump the hydrogen ions across the inner mitochondrial membrane
and create a proton gradient as evidenced in the low H+ concentration in the intermembrane.
Because of this, very little if any ATP is produced when the protons flow back across the
membrane through ATP synthase, which would contribute to Jared’s symptoms of muscle
weakness, shortness of breath, and dizziness.
Data
Explain how the data from your experiments showed that cyanide was used to poison Jared, and
that 2-deoxyglucose, arsenic and oligomycin were not used.
The data showed that cyanide poisoned Jared rather than the other 3 poisons as the relative changes
in pyruvate, NADH, and intermembrane H+ concentrations in Jared’s muscle cells in comparison
to the control (healthy) muscle cells of no change, increase, and decrease (respectively) only
matched the changes seen in the cyanide trial. Specifically, 2-deoxyglucose created a decreased in
pyruvate concentration, arsenic caused pyruvate concentration to increase and NADH
concentration to decrease, and oligomycin created an H+ increase as seen in the data from the
experiment step of the experiment. As such, only cyanide’s results paralleled the observations seen
in Jared’s muscle cells so the scientist could determine that cyanide was used to poison Jared.
Poisons
For each molecule of glucose, glycolysis only makes 2 ATP, the Krebs cycle only makes 2 ATP
and the ETC makes 34 ATP. If the suspects had given Jared the 2-deoxyglucose poison, would the
decrease in ATP concentration be smaller, the same or larger as when they used cyanide? Explain
your answer.
The decrease in ATP concentration would be larger than if they used cyanide as with cyanide, only
the final step of ATP production would be hampered in that the ETC is produce the 34 ATP from
the proton movement through the ATP synthase. However, since 2-deoxyglucose attaches to the
first enzyme of glycolysis, it halts the process and prevents the production of the 2 pyruvate
molecules, which creates a loss of the 2 ATP released from glycolysis and Krebs cycle in addition
to the 34 ATP of the ECT as without pyruvate and thus the NADH and FADH2 of the Krebs cycle,
the ECT cannot perform its function. In fact, since ATP must be inputted into glycolysis for the
first enzyme to operate, there would be a net loss of 34 + 2 + 2 + 1 = 39 ATP with 2-deoxyglucose
poison versus the 34 ATP of cyanide so the decrease in ATP concentration would be much larger.
Conclusions
Describe how the antidote helped Jared's cells make ATP (include the ETC and H+ gradient in
your answer).
The antidote helped Jared’s cells make ATP again as the antidote changed cyanide into thiocyanate
which the body can remove through urine. With the removal of cyanide from the ETC, the ETC
can pump a sufficient number of protons across the H+ gradient which can later flow back through
ATP synthase to create ATP once again.

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