A Genetic Code Expansion-Derived Molecular Beacon for the Detection of Intracellular Amyloid-β Peptide Generation

Angew Chem Int Ed Engl. 2021 Feb 19;60(8):3934-3939. doi: 10.1002/anie.202010703. Epub 2020 Dec 15.

Abstract

Polypeptides generated from proteolytic processing of protein precursors, or proteolytic proteoforms, play an important role in diverse biological functions and diseases. However, their often-small size and intricate post-translational biogenesis preclude the use of simple genetic tagging in their cellular studies. Herein, we develop a labeling strategy for this class of proteoforms, based on residue-specific genetic code expansion labeling with a molecular beacon design. We demonstrate the utility of such a design by creating a molecular beacon reporter to detect amyloid-β peptides, known to be involved in the pathogenesis of Alzheimer's disease, as they are produced from amyloid precursor protein (APP) along the endocytic pathway of living cells.

Keywords: amyloids; genetic code expansion; proteoforms; proteolytic processing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acyl-tRNA Synthetases / genetics
  • Amino Acyl-tRNA Synthetases / metabolism
  • Amyloid beta-Peptides / chemistry
  • Amyloid beta-Peptides / metabolism*
  • Amyloid beta-Protein Precursor / genetics
  • Amyloid beta-Protein Precursor / metabolism*
  • Archaeal Proteins / genetics
  • Archaeal Proteins / metabolism
  • Genetic Code
  • HEK293 Cells
  • Humans
  • Lysine / analogs & derivatives*
  • Lysine / chemistry
  • Lysine / metabolism
  • Methanosarcina / enzymology
  • Microscopy, Fluorescence
  • Mutagenesis, Site-Directed
  • Protein Processing, Post-Translational

Substances

  • Amyloid beta-Peptides
  • Amyloid beta-Protein Precursor
  • Archaeal Proteins
  • Amino Acyl-tRNA Synthetases
  • pyrrolysine
  • Lysine