Escherichia coli 70 S ribosome at 15 Å resolution by cryo-electron microscopy:: Localization of fMet-tRNAfMet and fitting of L1 protein

Cryo-electron microscopy of the ribosome in different binding states with mRNA and tRNA helps unravel the different steps of protein synthesis. Using over 29,000 projections of a ribosome complex in single-particle form, a three-dimensional map of the Escherichia coli 70 S ribosome was obtained in which a single site, the P site, is occupied by fMet-tRNA(f)(Met) as directed by an AUG codon containing mRNA. The superior resolution of this three-dimensional map, 14.9 Angstrom, has made it possible to fit the tRNA X-ray crystal structure directly and unambiguously into the electron density, thus determining the locations of anticodon-codon interaction and peptidyltransferase center of the ribosome. Furthermore, at this resolution, one of the distinctly visible domains corresponding to a ribosomal protein, L1, closely matches with its X-ray structure. (C) 1998 Academic Press.