Initiation of Protein Synthesis in HeLa Cells

Abstract

Initiation of protein synthesis in HeLa cells has been synchronized by exposure of the cells to fluoride. Double-labeling of such cells for short pulses with [³⁵S]methionine and a tritiated amino acid, followed by Edman degradation of the puromycin-released nascent peptides, has shown that the percent of N-terminal methionine incorporated compared to total incorporation is significantly higher than the value obtained with any of the other amino acids tested. The results suggest that the bulk of the nascent proteins synthesized in vivo by HeLa cells are initiated with methionine.

• synchronization of initiation
• Edman degradation
• N-terminal amino acids