Factors affecting protein synthesis in vitro in rabbit reticulocytes

Abstract

Rabbit reticulocytes in vitro rapidly incorporate labeled amino acids into their proteins. The process is accelerated by the plasma of every mammal investigated and also by extracts of normal erythrocytes, rabbit reticulocytes, liver, spleen, and yeast (1). We have described two sets of stimulating factors: one of these sets consists of certain amino acids (1), the other of fructose-amino acids in liver (2-4). The latter set is ineffective without the addition of iron to the reaction medium. The effect of iron has been referred to in preceding publications (2-5), but without detail. After the necessity of adding iron was recognized, in order to obtain a maximal rate of protein synthesis the reaction mixture was improved further by adding to it certain substances which depend upon added iron for their effect. These increased the effect of plasma. Eventually the total (potential as well as actual) accelerating effects of plasma and liver extract were accounted for by known substances. This led to the devising of a reaction mixture formula in which the amino acid incorporation is about five times as fast as that observed when the cells are incubated in saline