You are the senior biochemist for NextGenSugar Inc., who make artificial sugars.

Question

You are the senior biochemist for NextGenSugar Inc., who make artificial sugars. They have generated several sugar analogs, but they don’t know anything about their metabolism. You are tasked with determining how these analogs will contribute to metabolism and whether they will kill people.

At your disposal is radioactive labeled (radio-labeled) sugar analogs, mammalian cells that grow in growth medium that you can supplement with these sugars and incubators that allow you to control oxygen availability. The first class of compounds you distinguish are those that can enter cells and those that cannot. This is based on incubating cells with sugar analogs and seeing if the cells picked up radioactivity; no radioactivity meant that particular analog was not a substrate for GLUT1 transporter.

Using the following data, determine which enzymes were unable to utilize the particular sugar analogs and the step of the pathway(s) that is inhibited.

1:

After setting the oxygen levels to 0 in the incubator, you incubate the cells with media containing only individual sugar analogs alone. Of the 55 compounds tested 30 result in all the cells dying. The remaining 25 successfully went through which pathway?

If these 25 analogs were radio-labeled, what radio-labeled chemical would be found in the media?

2:

Of the 30 compounds that resulted in cell death, 1 was incubated with glucose in the media and caused increased consumption of the glucose, though still little metabolism of the analog. Describe a scenario where this would be the result. Which enzyme is this sugar analog a substrate for and which is it not?

3:            You now test the 25 compounds that allowed survival without oxygen in a full oxygen environment. 1 compound, while allowing for full ATP synthesis and cell survival, never allowed cells to divide or grow. What pathway was this analog incapable of participating in. Briefly describe the biochemical and cellular consequences that arose in this situation that led to the observed phenotypes (survival w/o division or growth).

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Explanation / Answer

1. Out of the 55 cells, only 25 could grow in an environment without oxygen. This implies , that these cells could use the sugar without the presence of oxygen, thus it could be said that these organisms were using sugar anaerobically. The cells went through an anaerobic glycolysis where the sugars would be metabolised in oxygen starved conditions. These sugars are capable of undergoing an anaerobic metabolism, similar to glucose. The radio labeled chemicals that would be found in the media are the different sugar analogs which cannot be specified here as there are several sugar anologues in this case; specifically 25 different analogues .

2. Of the 30 compounds that resulted in cell death, 1 was incubated with glucose in the media and caused increased consumption of the glucose, though still little metabolism of the analog. This would happen because the analog might not have been able to convert itself into glucose for further metabolism under oxygen starved condition. As a result, the glucose in the media would be consumed more easily, even under oxygen starved condition as glucose can undergo an anaerobic glycolysis , which the analog might not be capable of .

This enzyme would probably be a substrate for a hexokinase, if not particularly glucokinase, as hexose sugar would be metabolised by the cells .

3. The sugar metabolism is involved in cell proliferation as it provides high amount of energy plus carbon sources to aid in biomass production. Though the cells have been able to survive, in order to proliferate, the cells need to undergo lactic acid fermentation, that would occur immediately following the sugar metabolism that provides extra carbon for cell biomass growth . The sugar would probably not be able to undergo this pathway in order to allow cell division.

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