Transfer of reducing equivalents across the mitochondrial membrane. I. Hydrogen transfer mechanisms involved in the reduction of pyruvate to lactate in isolated liver cells.
(1) The reduction of pyruvate to lactate has been studied in isolated liver cells in order to elucidate the mechanims involved in the transfer of reducing equivalents from mitochondria to cytosol. (2) Manipulation of the cytosolic oxaloacetate concentration did not support the malate-oxaloacetate cycle as being responsible for the transfer of reducing equivalents out of the mitochondria: (a) With pyruvate plus oleate present 2 mM Amytal caused a 10-fold decrease in the oxaloacetate concentration, but had only a small inhibitory effect on lactate production. Oleate was essential in order to prevent disintegration of the cells in the presence of Amytal. (b) Quinolinate, an inhibitor of phosphoenolpyruvate carboxylase (GTP: oxaloacetate carboxylyase, transphosphorylating, EC 4.1.1.32), caused a several-fold increase in the oxaloacetate concentration but inhibited lactate production from pyruvate; this was accompanied by an increased reduction of mitochondrial pyridine nucleotides. (3) p-Chlorophenyl pyruvate, an inhibitor of pyruvate carboxylase (pyruvate: carbondioxide ligase, ADP, EC 6.4.1.1), also inhibited lactate production from pyruvate. (4) It is postulated that with pyruvate as substrate, recycling of carbon via pyruvate carboxylase, phosphoenolpyruvate carboxylase and pyruvate kinase (ATP: pyruvate phosphotransferase, EC 2.7.1.40) is an important, energy-requiring, mechanism for the transfer of the proportion of NADH not directly associated with gluconeogenesis.
Meijer
A J
AJ
Johnson Research Foundation, University of Pennsylvania, Philadelphia, PA 19174, USA.
Williamson
J R
JR
D000654:Amobarbital / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D005943:Gluconeogenesis / Q000502:physiology; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006859:Hydrogen / Q000378:metabolism*; D006993:Hypnotics and Sedatives / Q000494:pharmacology; D019344:Lactic Acid / Q000378:metabolism*; D008297:Male; D051336:Mitochondrial Membranes / Q000187:drug effects / Q000378:metabolism*; D019301:Oleic Acid / Q000378:metabolism; D010084:Oxidation-Reduction; D019289:Pyruvic Acid / Q000737:chemistry / Q000378:metabolism*; D017378:Quinolinic Acid / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D006993:Hypnotics and Sedatives; D019301:Oleic Acid; D019344:Lactic Acid; D006859:Hydrogen; D019289:Pyruvic Acid; D017378:Quinolinic Acid; D000654:Amobarbital