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Reaction 3: The Second Investment of ATP
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Description
Phosphofructokinase catalyzes a second ATP-dependent phosphorylation, to give a hexose derivative, fructose-1,6-bisphosphate (FBP), phosphorylated at both carbons 1 and 6.

Transferring the carbonyl oxygen from carbon 1 to carbon 2 has two important effects:

The hydroxyl group generated at carbon 1 can be readily phosphorylated in the reaction 3.

It also sets up the sugar for a symmetric aldol cleavage in reaction 4.

The phosphofructokinase reaction is the primary step at which glycolysis is regulated.
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