Insulin does not change the intracellular distribution of hexokinase in rat heart.
Preliminary evidence has suggested that hexokinase in rat heart changes its kinetic properties in response to insulin through translocation to the outer mitochondrial membrane. We reexamined this hypothesis in light of tracer kinetic evidence to the contrary. Our methods were as follows. Working rat hearts were perfused with Krebs-Henseleit buffer containing glucose (5 mmol/l) and sodium oleate (0.4 mmol/l). Dynamic glucose uptake was measured with [2-3H]glucose and with 2-deoxy-2-[18F]fluoroglucose (2-[18F]DG). Hexokinase activity was determined in the cytosolic and mitochondrial fractions. Our results are as follows. Uptake of glucose and uptake of 2-[18F]DG were parallel. Insulin (1 mU/ml) increased glucose uptake threefold but had no effect on 2-[18F]DG uptake. The tracer-to-tracee ratio decreased significantly. The Michaelis-Menten constant of hexokinase for 2-deoxyglucose was up to 10 times higher than for glucose. There was no difference in maximal reaction velocity. Two-thirds of hexokinase was bound to mitochondria. Insulin neither caused translocation nor changed Michaelis-Menten constant or maximal reaction velocity. In conclusion, the insulin-induced changes in the tracer-to-tracee ratio are due to a shift of the rate-limiting step for glucose uptake from transport to phosphorylation by hexokinase. Insulin does not affect the intracellular distribution or the kinetics of hexokinase.
Doenst
T
T
Department of Medicine, Division of Cardiology, University of Texas-Houston Medical School, Houston, Texas 77030, USA.
Han
Q
Q
Goodwin
G W
GW
Guthrie
P H
PH
Taegtmeyer
H
H
D000818:Animals; D001692:Biological Transport; D002950:Citrate (si)-Synthase / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D005215:Fasting; D019788:Fluorodeoxyglucose F18 / Q000493:pharmacokinetics*; D005947:Glucose / Q000378:metabolism*; D006321:Heart / Q000187:drug effects* / Q000502:physiology; D006593:Hexokinase / Q000378:metabolism*; D066298:In Vitro Techniques; D007328:Insulin / Q000494:pharmacology* / Q000502:physiology; D007700:Kinetics; D008297:Male; D008929:Mitochondria, Heart / Q000201:enzymology*; D009200:Myocardial Contraction; D009206:Myocardium / Q000201:enzymology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014316:Tritium
D007328:Insulin; D019788:Fluorodeoxyglucose F18; D014316:Tritium; D002950:Citrate (si)-Synthase; D006593:Hexokinase; D005947:Glucose