Small Unnatural Amino Acid Carried Raman Tag for Molecular Imaging of Genetically Targeted Proteins

J Phys Chem Lett. 2018 Aug 16;9(16):4679-4685. doi: 10.1021/acs.jpclett.8b01991. Epub 2018 Aug 3.

Abstract

Raman has been implemented to image biological systems for decades. However, Raman microscopy along with Raman probes is restricted to image metabolites or a few intracellular organelles so far and lacks genetic specificity for imaging proteins of interest, which significantly hinders their application. Here, we report the Raman spectra-based protein imaging method, which incorporates a small phenyl ring enhanced Raman tag (total of ∼0.55 kDa) with a single unnatural amino acid (UAA) to genetically label specific proteins. We further demonstrate hyperspectral stimulated Raman scattering (SRS) imaging of the Histone3.3 protein in the nucleus, Sec61β protein in the endoplasmic reticulum of HeLa cells, and Huntingtin protein Htt74Q in mutant huntingtin-induced cells. Genetic encoding of a small, stable, sensitive, and narrow-band Raman tag took one key step forward to enable SRS or Raman imaging of specific proteins and could further facilitate quantitative Raman spectra-based supermultiplexing microscopy in the future.

Publication types

  • Letter

MeSH terms

  • Alkynes / chemical synthesis
  • Alkynes / chemistry*
  • Benzamides / chemical synthesis
  • Benzamides / chemistry*
  • HeLa Cells
  • Histones / genetics
  • Histones / metabolism
  • Humans
  • Huntingtin Protein / genetics
  • Huntingtin Protein / metabolism
  • Lysine / analogs & derivatives*
  • Lysine / chemical synthesis
  • Lysine / chemistry*
  • Lysine / genetics
  • Molecular Imaging / methods*
  • Molecular Probes / chemical synthesis
  • Molecular Probes / chemistry*
  • Molecular Probes / genetics
  • Proteins / genetics
  • Proteins / metabolism*
  • SEC Translocation Channels / genetics
  • SEC Translocation Channels / metabolism
  • Spectrum Analysis, Raman / methods

Substances

  • Alkynes
  • Benzamides
  • HTT protein, human
  • Histones
  • Huntingtin Protein
  • Molecular Probes
  • Proteins
  • SEC Translocation Channels
  • SEC61B protein, human
  • Lysine