NADP-malate dehydrogenase activity in rat erythrocytes. Comparison with pyruvate kinase in relation to coupling with lactate dehydrogenase.
The present study explores the possible channelling of pyruvate generated by either pyruvate kinase or NADP-malate dehydrogenase to lactate dehydrogenase in cross-linked and permeabilized erythrocytes. The generation of both unlabelled and 14C-labelled pyruvate and lactate was measured in rat erythrocytes, which were prepared for cross-linking with dimethyl suberimidate and permeabilization by digitonin and then exposed to unlabelled or 14C-labelled malate and/or phospho-enol-pyruvate. Rat erythrocytes were found to display NADP-malate dehydrogenase activity. Under conditions in which the generation rates of pyruvate from either phospho-enol-pyruvate (15 microM) or malate (0.5 mM) were not vastly different from one another, a greater fraction of the 2-keto acid was converted to lactate when produced from phospho-enol-[1-14C]pyruvate rather than [U-14C]malate. This difference was most obvious when the availability of exogenous NADH was close to or somewhat below that theoretically required to ensure full conversion of endogenously formed pyruvate to lactate. These findings are compatible with the view that pyruvate generated at the pyruvate kinase level is converted to lactate more efficiently than pyruvate produced in the reaction catalysed by NADP-malate dehydrogenase.
Malaisse-Lagae
F
F
Laboratory of Experimental Medicine, Erasmus Medical School, Brussels Free University, Belgium.
Zähner
D
D
Malaisse
W J
WJ
D000818:Animals; D002478:Cells, Cultured; D004912:Erythrocytes / Q000201:enzymology*; D007515:Islets of Langerhans / Q000201:enzymology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D007773:Lactates / Q000378:metabolism; D019344:Lactic Acid; D008291:Malate Dehydrogenase / Q000097:blood*; D050538:Malate Dehydrogenase (NADP+); D011770:Pyruvate Kinase / Q000378:metabolism*; D051381:Rats
D007773:Lactates; D019344:Lactic Acid; D007770:L-Lactate Dehydrogenase; D008291:Malate Dehydrogenase; D050538:Malate Dehydrogenase (NADP+); D011770:Pyruvate Kinase