Sensitive Separation Procedure for Escherichia coli Ribosomal Proteins and the Resolution of High-Molecular-Weight Components

Abstract

Application of the two-dimensional polyacrylamide gel electrophoretic procedure recently described by Mets and Bogorad [4] to the resolution of Escherichia coli ribosomal proteins is described. This procedure is fast (running time of 8–10 h) and highly sensitive (1–2 μg protein per spot) and it gave good resolution for the majority of the 55 proteins in E. coli ribosomes. Moreover, the method has permitted the resolution of a large number of high-molecular-weight proteins associated with ribosomes not exposed to high-salt washing. Using purified proteins, the positions in the electrophrogram of the individual proteins of the 30–S subunit have been determined.