Marketing and the U. S. Supreme Court, 1968-1974

Immunological Techniques for Studies on the Biogenesis of Mitochondrial Membrane Proteins

“…In the ca se of cytochrome oxidase (Schwab et al , 1972;Sebald et al, 1973;Werner, 1974), cytochrome b (Weiss and Ziganke, 1974;Weiss, 1976), and the ATPase complex (Se bald, 1977), 60 to 120 min are necessary until all radioactivity appears in the "mature" protein.…”

“…A continuous labeling of all cellular proteins, that is, precursors and mature proteins, is obtained when the radio active amino acid is applied in high amounts (Werner, 1974). About 300 /Lmole of leucine are required for the synthesis of 500 mg of cellular protein.…”

“…This type of labeling alJows the direct evaluation of protein synthesis rates as weil as the amount of precursors present in the celJ (Werner, 1974;Werner et al, 1974). [35S] sulfate.…”

“…The Triton concentration may be varied (depending on the amount of mitochondrial protein) between 0.5 and 3% without interfering with the subsequent immunoprecipitation. The lysate is centrifuged for 20 min at 50,000 x g. Ninety-five percent of the total mitochondrial protein is found in the supernatant (Werner 1974) if the procedure was performed with freshly prepared mitochondria. For mitochondrial suspensions which have been frozen and thawed repeatedly prior to the addition of Triton X-loO, the solubilization rate may drop to 70%.…”

“…The aim of the assays was to characterize two antibody preparations obtained from rabbits: immunoglobulin to cytochrome oxidase from Neurospora crassa and immunoglobulin to the isolated subunit 3 of the same enzyme (Werner, 1974). The reactions of the antibodies to the purified membrane components and to whole mitochondrial membrane extracts were examined.…”