Transport and metabolism of L-glutamate during oxygenation, anoxia, and reoxygenation of rat cardiac myocytes.
The intracellular glutamate concentration of oxygenated, isolated adult rat heart cells incubated with 0.15 mM glutamate amounts to 2.89 +/- 0.6 mM. Under these conditions the velocity of glutamate transport was 24.3 +/- 1.6 pmol.min-1.mg protein-1 and occurs via a high-affinity carrier characterized by an apparent affinity (K(m)) value of 0.18 +/- 0.03 mM. At high glutamate concentrations ( > 1mM) this high-affinity transport system is superimposed by additional uptake processes of a low affinity but a high capacity for glutamate. The 1.6-fold increased uptake of glutamate observed during 30 min of anoxic incubation of cardiomyocytes does not prevent an intracellular decrease in this amino acid to a concentration of 0.49 mM. After 15 min reoxygenation of cardiomyocytes the intracellular glutamate content increases to the control values of oxygenated cells. Only 2.4% of the glutamate increase after reoxygenation is due to the transport o glutamate from the incubation medium. The competitive inhibitor of transaminases, aminooxyacetate, prevents both the observed intracellular decrease in glutamate during anoxia and the increase in intracellular glutamate after reoxygenation of cardiomyocytes. Half of the amino groups needed for the synthesis of glutamate originate from intracellular alanine, which increases during anoxia and is metabolized during reoxygenation of cardiomyocytes. The velocity of the glutamate uptake of cardiomyocytes incubated in a medium containing 10 mM L-glutamate amounted to 728 +/- 140 pmol.min-1.mg protein-1. During anoxic incubation of cardiomyocytes at this high extracellular glutamate concentration, the intracellular glutamate breakdown may be compensated by a simultaneous uptake of this amino acid via the transport processes characterized by a high capacity
Dinkelborg
L M
LM
Institut für Zoophysiologie, Lehrstuhl für Stoffwechselphysiologie, Heinrich-Heine-Universität, Düsseldorf, Germany.
Kinne
R K
RK
Grieshaber
M K
MK
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001692:Biological Transport; D004734:Energy Metabolism; D018698:Glutamic Acid / Q000493:pharmacokinetics*; D006321:Heart / Q000187:drug effects*; D000860:Hypoxia / Q000378:metabolism* / Q000473:pathology; D007425:Intracellular Membranes / Q000378:metabolism; D019344:Lactic Acid / Q000096:biosynthesis; D008297:Male; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D010100:Oxygen / Q000008:administration & dosage*; D010710:Phosphates / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012508:Sarcolemma / Q000378:metabolism
D000596:Amino Acids; D010710:Phosphates; D019344:Lactic Acid; D018698:Glutamic Acid; D010100:Oxygen