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

J Mol Biol. 1998 Jul 3;280(1):103-16. doi: 10.1006/jmbi.1998.1859.

Abstract

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-tRNAfMet as directed by an AUG codon containing mRNA. The superior resolution of this three-dimensional map, 14.9 A, 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.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism
  • Binding Sites
  • Cryoultramicrotomy
  • Crystallography, X-Ray
  • Escherichia coli / genetics
  • Image Processing, Computer-Assisted
  • Microscopy, Electron
  • Models, Molecular
  • Nucleic Acid Conformation*
  • Peptides
  • Protein Conformation*
  • RNA, Transfer / metabolism
  • RNA, Transfer, Met / chemistry
  • RNA, Transfer, Met / metabolism
  • RNA, Transfer, Met / ultrastructure*
  • Ribosomal Proteins / chemistry*
  • Ribosomal Proteins / metabolism
  • Ribosomes / metabolism
  • Ribosomes / ultrastructure*
  • Spectroscopy, Fourier Transform Infrared / methods
  • Thermus thermophilus / metabolism

Substances

  • Bacterial Proteins
  • Peptides
  • RNA, Transfer, Met
  • Ribosomal Proteins
  • fMet-tRNA(fMet)
  • ribosomal protein L1
  • RNA, Transfer