Abstract
Midgut mitochondria from fifth larval instar Manduca sexta exhibit a membrane-associated transhydrogenase that catalyzes hydride ion transfer between NADP(H) and NAD(H). The NADPH-forming transhydrogenations occur as nonenergy- and energy-linked activities. The energy-linked activities couple with electron transport-dependent utilization of NADH/succinate, or with Mg2+-dependent ATPase. These energy-linked transhydrogenations have been shown to be physiologically and developmentally significant with respect to insect larval/pupal maturation. In the present study, isolated mitochondrial membranes were lyophilized and subjected to organic solvent or phospholipase treatments. Acetone extraction and addition of Phospholipase A2 proved to be effective inhibitors of the insect transhydrogenase. Liberation of phospholipids was reflected by measured phosphorous release. Addition of phospholipids to organic solvent- and phospholipase-treated membranes was without effect. Employing a partially lipid-depleted preparation, phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine were reintroduced and transhydrogenase activity assessed. Of the phospholipids tested, only phosphatidylcholine significantly stimulated transhydrogenase activity. The results of this study suggest a phospholipid dependence of the M. sexta mitochondrial transhydrogenase.
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This study was supported in part by Houghton College and the United States Army Reserve, APMC (KPV). The assistance of Dr. Martin Mitchell, Edinboro University of Pennsylvania, for his review is gratefully acknowledged.
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Vandock, K.P., Emerson, D.J., McLendon, K.E. et al. Phospholipid Dependence of the Reversible, Energy-Linked, Mitochondrial Transhydrogenase in Manduca sexta . J Membrane Biol 242, 89–94 (2011). https://doi.org/10.1007/s00232-011-9379-1
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DOI: https://doi.org/10.1007/s00232-011-9379-1