Dear all,
Apologies for a off topic question. I could not think of a better
place to ask this.
Suppose, we have four enzymes turning over five molecules, either in
the living cell, or in fresh mammalian cell lyset. The molecules could
be as simple as ATP, GTP, NADP, GSH etc. Now, one can add a inhibitor
/ drug Y to this mixture and that might change the dynamics. I am
wondering how could someone measure the concentration of these
metabolites.. ! It can not be done on purified enzyme, it has to be
done when everything else is present.
All I can think of are:
a) Using mass spec on the metabolites: Quench the reaction at
different time points, remove whatever possible by centrifugation and
a spin filter (similar to QIAGEN kit), and do mass spec.
b) Use some sort of a focused optical method (IR/UV laser?) on living
cell that could excite multiple molecules (but I have no clue beyond
this..)
Has anybody seen something that addresses the question? Any suggestion
would be highly appreciated.
Cheers, Partha
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