Evidence of a novel role for monovalent cations in pyruvate kinase catalysis.
In the pyruvate kinase (EC 2.7.1.40) reaction, the complete enzyme-products complex consists of enzyme, pyruvate, ATP, a divalent cation, and a monovalent cation, usually K+. The dissociation of this complex can be slow permitting reversible enolization of pyruvate in this complex during the course of the forward reaction. High concentrations of each component in the enzyme-products complex inhibits the forward reaction, but only elevated concentrations of the monovalent cation decrease the net rate of product dissociation relative to that of pyruvate enolization. This result indicates that the monovalent cation can be the first component released from the enzyme-products complex and that the presence of an inhibiting monovalent cation concentration leads to re-formation of the complete complex, which is necessary for pyruvate enolization. The evidence suggests that the monovalent cation can bind and release with each turnover of the enzyme. While the data do not permit the conclusion that first release of monovalent cation is the exclusive pathway for dissociation of the enzyme-products complex, no other component once released can reassociate rapidly enough to form a complete complex during the forward reaction. Inhibition by these components must be attributed to the formation of abortive complexes.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D002414:Cations, Monovalent; D003035:Cobalt / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D009132:Muscles / Q000201:enzymology; D009994:Osmolar Concentration; D011188:Potassium / Q000494:pharmacology; D011485:Protein Binding; D011770:Pyruvate Kinase / Q000378:metabolism*; D011817:Rabbits
D002414:Cations, Monovalent; D000643:Ammonium Chloride; D003035:Cobalt; D000255:Adenosine Triphosphate; D011770:Pyruvate Kinase; D008274:Magnesium; D011188:Potassium