Incorporation in vitro of [14C]Leucine into the Mitochondrial Protein of Lucilia cuprina

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Abstract
The energy requirements for incorporation in vitro of [14C]leucine into sterile mitochondria isolated from thoracic flight muscles of newly‐emerged Lucilia cuprina have been studied.There is an absolute requirement for addition of either ADP or ATP. Oxidisable substrates (pyruvate, DL‐α‐glycerophosphate, proline), or an ATP‐generating system increased incorporation above that observed with added ADP or ATP alone. Both the rate and duration of incorporation are strongly dependent on the concentration of adenine nucleotides used. Experiments with oligomycin and 2,4‐dinitrophenol suggest that energy for incorporation is provided as ATP.With non‐sterile preparations, incorporation was, independent of added adenine nucleotides, and almost constant for up to four hours with no added substrate or adenine nucleotide. It was inhibited by added substrate and also by added amino acids, especially isoleucine and valine.