Semi-synthesis of labeled proteins for spectroscopic applications

Molecules. 2013 Jan 2;18(1):440-65. doi: 10.3390/molecules18010440.

Abstract

Since the introduction of SPPS by Merrifield in the 60s, peptide chemists have considered the possibility of preparing large proteins. The introduction of native chemical ligation in the 90s and then of expressed protein ligation have opened the way to the preparation of synthetic proteins without size limitations. This review focuses on semi-synthetic strategies useful to prepare proteins decorated with spectroscopic probes, like fluorescent labels and stable isotopes, and their biophysical applications. We show that expressed protein ligation, combining the advantages of organic chemistry with the easy and size limitless recombinant protein expression, is an excellent strategy for the chemical synthesis of labeled proteins, enabling a single protein to be functionalized at one or even more distinct positions with different probes.

Publication types

  • Review

MeSH terms

  • Amino Acid Motifs
  • Fluorescence Resonance Energy Transfer
  • Fluorescent Dyes / chemistry*
  • Humans
  • Inteins
  • Isotope Labeling
  • Nuclear Magnetic Resonance, Biomolecular
  • Peptide Fragments / biosynthesis
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Protein Engineering
  • Proteins / chemistry*
  • Proteins / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Staining and Labeling / methods

Substances

  • Fluorescent Dyes
  • Peptide Fragments
  • Proteins
  • Recombinant Proteins