Design, Synthesis, and Evaluation of N- and C-Terminal Protein Bioconjugates as G Protein-Coupled Receptor Agonists

Bioconjug Chem. 2018 Feb 21;29(2):403-409. doi: 10.1021/acs.bioconjchem.7b00716. Epub 2018 Feb 2.

Abstract

A G protein-coupled receptor (GPCR) agonist protein, thaumatin, was site-specifically conjugated at the N- or C-terminus with a fluorophore for visualization of GPCR:agonist interactions. The N-terminus was specifically conjugated using a synthetic 2-pyridinecarboxyaldehyde reagent. The interaction profiles observed for N- and C-terminal conjugates were varied; N-terminal conjugates interacted very weakly with the GPCR of interest, whereas C-terminal conjugates bound to the receptor. These chemical biology tools allow interactions of therapeutic proteins:GPCR to be monitored and visualized. The methodology used for site-specific bioconjugation represents an advance in application of 2-pyridinecarboxyaldehydes for N-terminal specific bioconjugations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Drug Design
  • Fluorescence
  • Fluorescent Dyes / chemistry
  • Humans
  • Microscopy, Fluorescence / methods
  • Optical Imaging
  • Plant Proteins / chemistry*
  • Plant Proteins / pharmacology*
  • Protein Binding
  • Receptors, G-Protein-Coupled / agonists*
  • Receptors, G-Protein-Coupled / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / pharmacology
  • Sweetening Agents / chemistry*
  • Sweetening Agents / pharmacology*

Substances

  • Fluorescent Dyes
  • Plant Proteins
  • Receptors, G-Protein-Coupled
  • Recombinant Proteins
  • Sweetening Agents
  • thaumatin protein, plant