Abstract
The effect of ionic strength during isolation on the activity of chicken liver ribosomes in a poly(U)-dependent phenylalanine-incorporating system was studied. Changing KCl concentration during isolation caused ribosome yield and activity to vary significantly. KCl concentrations of 150–250 mM in the isolation medium resulted in higher and more consistent ribosome yields and activity than 25–50 mM, although KCl concentration was the same in all assay mixtures (150 mM). Thus, the reduced activity characteristic of chicken ribosomes isolated at low KCl concentrations is not readily reversible in the assay mixture. The reduced yield and activity may result from a greater tendency of ribosomes isolated at low KCl concentrations to aggregate. The effects of cell-sap concentration, MgCl 2 concentration, and temperature during incubation were also determined, as well as the results of freezing ribosomes and cell sap before use. The optimum MgCl 2 concentration for phenylalanine incorporation was about 14 mM, and the optimum temperature for a 60-min incubation was 33–37 °C.
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