Phosphorylation of glycolytic and gluconeogenic enzymes by the insulin receptor kinase.
Various glycolytic and gluconeogenic enzymes were tested as substrates for the insulin receptor kinase. Phosphofructokinase and phosphoglycerate mutase were found to be the best substrates. Phosphorylation of these enzymes was rapid, stimulated 2- to 6-fold by 10(-7) M insulin and occurred exclusively on tyrosine residues. Enolase, fructose 1,6-bisphosphatase, lactate dehydrogenases in decreasing order, were also subject to insulin-stimulated phosphorylation but to a smaller extent than that for phosphofructokinase or phosphoglycerate mutase. The phosphorylation of phosphofructokinase was studied most extensively since phosphofructokinase is known to catalyze a rate-limiting step in glycolysis. The apparent Km of the insulin receptor for phosphofructokinase was 0.1 microM, which is within the physiologic range of concentration of this enzyme in most cells. Tyrosine phosphorylation of phosphofructokinase paralleled autophosphorylation of the beta-subunit of the insulin receptor with respect to time course, insulin dose response (half maximal effect between 10(-9) and 10(-8) M insulin), and cation requirement (Mn2+ greater than Mg2+ much greater than Ca2+). Further study will be required to determine whether the tyrosine phosphorylation of phosphofructokinase plays a role in insulin-stimulated increases in glycolytic flux.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001731:Bisphosphoglycerate Mutase / Q000378:metabolism; D002412:Cations / Q000494:pharmacology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D006597:Fructose-Bisphosphatase / Q000378:metabolism; D005943:Gluconeogenesis; D006019:Glycolysis; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D010732:Phosphofructokinase-1 / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D051381:Rats; D011972:Receptor, Insulin / Q000378:metabolism*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D002412:Cations; D007328:Insulin; D010750:Phosphoproteins; D019000:Phosphotyrosine; D014443:Tyrosine; D000255:Adenosine Triphosphate; D007770:L-Lactate Dehydrogenase; D010732:Phosphofructokinase-1; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin; D006597:Fructose-Bisphosphatase; D010751:Phosphopyruvate Hydratase; D001731:Bisphosphoglycerate Mutase