Affiliation:
1. Laboratoire de Chimie Physiologique, Université de Louvain and International Institute of Cellular and Molecular Pathology, U.C.L. 75.39, B-1200 Brussels, Belgium
Abstract
1. Recycling of metabolites between fructose 6-phosphate and triose phosphates has been investigated in isolated hepatocytes by the randomization of carbon between C(1) and C(6) of glucose formed from [1-14C]galactose. 2. Randomization of carbon atoms was regularly observed with hepatocytes isolated from fed rats and was then little influenced by the concentration of glucose in the incubation medium. It was decreased by about 50% in the presence of glucagon. 3. Randomization of carbon atoms by hepatocytes isolated from starved rats was barely detectable at physiological concentrations of glucose in the incubation medium, but was greatly increased with increasing glucose concentrations. It was nearly completely suppressed by glucagon. These large changes can be attributed to parallel variations in the activity of phosphofructokinase. 4. The main factors that appear to control the activity of phosphofructokinase under these experimental conditions are the concentration of fructose 6-phosphate, the concentration of fructose 1,6-bisphosphate and also the affinity of the enzyme for fructose 6-phosphate. 5. The affinity of phosphofructokinase for fructose 6-phosphate was diminished by incubation of the cells in the presence of glucagon and also by filtration of an extract of hepatocytes through Sephadex G-25 and by purification of the enzyme. When assayed at 0.25 or 0.5mm-fructose 6-phosphate, the activity of phosphofructokinase present in a liver Sephadex filtrate was increased by a low-molecular-weight effector, which could be isolated from a liver extract by ultrafiltration, gel filtration or heat treatment, but was rapidly destroyed in trichloroacetic acid, even in the cold. This effector appears to be a highly acid-labile phosphoric ester. Its concentration was greatly increased in hepatocytes incubated in the presence of glucose and was decreased in the presence of glucagon.
Subject
Cell Biology,Molecular Biology,Biochemistry
Cited by
204 articles.
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