Gene knockouts are a common method to determine the functions of unknown protein
ID: 3519571 • Letter: G
Question
Gene knockouts are a common method to determine the functions of unknown proteins. This method selectively prevents one particularly protein from being expressed in an organism. Biochemical methods are then used to determine the effect on each pathway Two enzymes from the metabolic pathways we've covered have been knocked out in different organisms Neither is able to reach maturity, but a cells were incubated and examined in cach case. In cach of the two organisms below, determine which enzyme was knocked out. Justify your answer in cach case by walking through your logic. Organism I: (15 points) Cells are grown in a rich source of glucose. Glutamate was included but proved toxic. Lactic acid concentration was increased. 2.4-DNP had little effect. While there was an increase in 5-ribose concentration, levels of nucleotides, specifically purines, were abnormally low without supplements. ATP levels were measured with the addition of various substrates: No addition Pyruvate Succinyl-CoA 6 NADH 10-12 10-12 20-22 25-27 Organism 2: (15 points) Cells are grown in a rich source of glucose. Lactic acid concentration was increased. 2,4-DNP had no effect. Cells showed a substantial improvement with the addition of glutamate. Levels of non-essential amino acids, lipids, and nucleotides was lower than anticipated and cells were very fragile without supplements. Plenty of 5-ribose was found in the cells. ATP levels were measured with the addition of various substrates: ximate No addition Pyruvate Succinyl-CoA 6 NADH 5-7 5-7 25-27 20-22Explanation / Answer
Organism 1:
As case is described, Addition of Glutamate has no effect and addition of Succinyle Co. A led to tremendous increase in ATP production, most probable enzyme knocket out was ?-Keto Glutarate Dehydrogenase enzyme. This led to shifting of pathway to production of Lactic Acid production, so addition of 2NADH led to increase in ATP production. So addition of Pyruvate had no effect.
Organism 1:
As case is described, Addition of Glutamate led to tremendous increase in ATP productionno effect and plenty of 5 ribose found in the cell, most probable enzyme knocket out was Pyruvate Dehydrogenase enzyme. This led to shifting of pathway to production of plenty of 5 ribose production, so addition of Succinyle Co. A led to tremendous increase in ATP production.
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