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Topic: How can you tell if O-18 has been incorporated into phosphate?  (Read 3817 times)

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Offline joyb

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The hydrolysis of D-glucose-1-phosphate to D-glucose and phosphate is catalysed by
alkaline phosphatase. If the enzyme catalysed reaction is conducted in H218O, 18O is incorporated into the phosphate product but is not incorporated into glucose. Suggest what experimental technique was used to demonstrate the incorporation of 18O into phosphate.

I'm stuck on this question- how can you specifically tell that the 18O has been incorporated into phosphate but not glucose?

Offline sjb

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Re: How can you tell if O-18 has been incorporated into phosphate?
« Reply #1 on: April 05, 2013, 07:30:57 AM »
The hydrolysis of D-glucose-1-phosphate to D-glucose and phosphate is catalysed by
alkaline phosphatase. If the enzyme catalysed reaction is conducted in H218O, 18O is incorporated into the phosphate product but is not incorporated into glucose. Suggest what experimental technique was used to demonstrate the incorporation of 18O into phosphate.

I'm stuck on this question- how can you specifically tell that the 18O has been incorporated into phosphate but not glucose?

How could you tell if it were incorporated into anything?

Offline Arkcon

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Re: How can you tell if O-18 has been incorporated into phosphate?
« Reply #2 on: April 05, 2013, 09:06:02 AM »
Like sjb: said, what is different about 18O?  How was anyone aware that it was even in the water used for the experiment?  This is, kinda, a very simple textbook definition.  Either you look it up, or you miss it.  We can't really "help" you find this out.  Unless you think about all the elements, that have isotopes, and how we'll tell them apart.
Hey, I'm not judging.  I just like to shoot straight.  I'm a man of science.

Offline Babcock_Hall

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Re: How can you tell if O-18 has been incorporated into phosphate?
« Reply #3 on: April 05, 2013, 09:11:48 AM »
There is an obvious answer to this question, but there is also a less obvious answer involving NMR.

Offline joyb

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Re: How can you tell if O-18 has been incorporated into phosphate?
« Reply #4 on: April 05, 2013, 01:29:22 PM »
Could you put the sample in a mass spectrometer and find that the mass of the phosphate has increased but not the glucose?

Offline Arkcon

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Re: How can you tell if O-18 has been incorporated into phosphate?
« Reply #5 on: April 05, 2013, 04:45:03 PM »
Could you put the sample in a mass spectrometer and find that the mass of the phosphate has increased but not the glucose?

That is the one I was hoping you'd come up with.  Think about Babcock_Hall:'s method now, how would that one work?
Hey, I'm not judging.  I just like to shoot straight.  I'm a man of science.

Offline Babcock_Hall

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Re: How can you tell if O-18 has been incorporated into phosphate?
« Reply #6 on: April 06, 2013, 09:49:14 AM »
I'll give you another hint.  O-18 is not itself NMR-active.

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