Oxidation of reduced cytosolic nicotinamide adenine dinucleotide by the malate-aspartate shuttle in the K-562 human leukemia cell line.
The activity of the malate-aspartate shuttle for the reoxidation of reduced cytosolic nicotinamide adenine dinucleotide (NADH) by mitochondria was studied in a line of human myeloid leukemia cells (K-562). The tumor cells showed mitochondrial reoxidation of cytosolic NADH, as evidenced by the accumulation of pyruvate, when incubated aerobically with L-lactate. The involvement of the respiratory chain in the reoxidation of cytosolic NADH was demonstrated by the action of rotenone, antimycin A, and oligomycin which strongly inhibited the formation of pyruvate from added L-lactate. Moreover, pyruvate production was greatly inhibited by the transaminase inhibitor, aminooxyacetate. Under glycolytic conditions, in the presence of aminooxyacetate, the rate of pyruvate production was also markedly inhibited, the rate of lactate accumulation was stimulated, and at 60 min the cytosolic NADH/nicotinamide adenine dinucleotide (NAD) ratio had increased progressively about 5-fold with respect to untreated cells. The maximal rate of the malate-aspartate shuttle has also been established by addition of arsenite to inhibit mitochondrial oxidation of the pyruvate formed from added L-lactate.
D000625:Aminooxyacetic Acid / Q000494:pharmacology; D001151:Arsenic / Q000494:pharmacology; D018053:Arsenites; D001219:Aspartate Aminotransferases / Q000037:antagonists & inhibitors; D001224:Aspartic Acid / Q000378:metabolism; D002478:Cells, Cultured; D004579:Electron Transport / Q000187:drug effects; D006801:Humans; D007773:Lactates / Q000378:metabolism; D007942:Leukemia, Experimental / Q000378:metabolism*; D008293:Malates / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D009243:NAD / Q000378:metabolism*; D010084:Oxidation-Reduction; D011773:Pyruvates / Q000378:metabolism
D018053:Arsenites; D007773:Lactates; D008293:Malates; D011773:Pyruvates; D009243:NAD; D000625:Aminooxyacetic Acid; D001224:Aspartic Acid; D001219:Aspartate Aminotransferases; C015001:arsenite; D001151:Arsenic