Effect of Polyamines on Globin Synthesis in a Rabbit Reticulocyte Polyamine-Free Protein Synthetic System1

Abstract

A polyamine-free protein synthetic system, established from components of rabbit reticulocytes, consisted of globin mRNA, salt-washed ribosomes, partially purified initiation factors, and pH 5 enzymes. Spermidine added to this system not only lowered the optimal magnesium concentration required for globin synthesis, but it also stimulated the globin synthesis 6- to 8-fold. The optimal spermidine concentration was 0.4 to 0.6 mM, a concentration similar to that in intact rabbit reticulocytes. The ratio of alpha to beta globin chains synthesized in the presence of spermidine and Mg2+ was approximately 1.0, while the ratio in the presence of only Mg2+ was approximately 1.5. Formation of methionyl puromycin was stimulated 4- to 6-fold by spermidine. This indicates that the stimulation is mainly at the level of initiation.