Synthesis of mitochondrial protein in the flight muscles of the Colorado beetle. Differentiation in vivo between extra- and intra-mitochondrial contributions to the amino acid incorporation into mitochondrial protein.

Abstract

By using cycloheximide, an inhibitor of cytoplasmic protein synthesis, conditions were investigated to estimate in vivo the extra- and intra-mitochondrial contributions to the synthesis of organelle protein in the flight muscles of Colorado beetles. With 4-day-old beetles about 15% of the [(14)C]leucine incorporation into mitochondrial protein is resistant to the action of cycloheximide. The incorporation into cytosol protein is inhibited by more than 99.5% with cycloheximide. During the first hour after precursor administration the incorporation into mitochondrial protein proceeds, in both the presence and the absence of cycloheximide, at a more-or-less linear rate with time. The cycloheximide-resistant amino acid incorporation is sensitive to the inhibitor of mitochondrial protein synthesis, chloramphenicol. The uncertainties inherent in the use of cycloheximide were discussed in arriving at the conclusion that about 15% of the mitochondrial protein is formed inside the organelle.