Protein synthesis by single ribosomes

RNA. 2003 Oct;9(10):1174-9. doi: 10.1261/rna.5800303.

Abstract

The ribosome is universally responsible for synthesizing proteins by translating the genetic code transcribed in mRNA into an amino acid sequence. Ribosomes use cellular accessory proteins, soluble transfer RNAs, and metabolic energy to accomplish the initiation, elongation, and termination of peptide synthesis. In translocating processively along the mRNA template during the elongation cycle, ribosomes act as supramolecular motors. Here we demonstrate that ribosomes adsorbed on a surface, as for mechanical or spectroscopic studies, are capable of polypeptide synthesis and that tethered particle analysis of fluorescent beads connected to ribosomes via polyuridylic acid can be used to estimate the rate of polyphenylalanine synthesis by individual ribosomes. This work opens the way for application of biophysical techniques, originally developed for the classical motor proteins, to the understanding of protein biosynthesis.

Publication types

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

MeSH terms

  • Escherichia coli / metabolism
  • Fluorescent Dyes / chemical synthesis
  • Microscopy, Atomic Force
  • Peptide Biosynthesis*
  • Peptide Chain Elongation, Translational*
  • Peptide Elongation Factors / metabolism
  • Peptide Fragments / analysis
  • Peptides / metabolism
  • Poly U / metabolism
  • Protein Biosynthesis*
  • RNA, Transfer, Amino Acyl
  • Ribosomes / metabolism*
  • Solubility

Substances

  • Fluorescent Dyes
  • Peptide Elongation Factors
  • Peptide Fragments
  • Peptides
  • RNA, Transfer, Amino Acyl
  • Poly U
  • polyphenylalanine