Label-Free Analysis with Multiple Parameters Separates G Protein-Coupled Receptor Signaling Pathways

Anal Chem. 2020 Nov 3;92(21):14509-14516. doi: 10.1021/acs.analchem.0c02652. Epub 2020 Oct 15.

Abstract

Real-time label-free techniques are used to profile G protein-coupled receptor (GPCR) signaling pathways in living cells. However, interpreting the label-free signal responses is challenging, and previously reported methods do not reliably separate pathways from each other. In this study, a continuous angular-scanning surface plasmon resonance (SPR) technique is utilized for measuring label-free GPCR signal profiles. We show how the continuous angular-scanning ability, measuring up to nine real-time label-free parameters simultaneously, results in more information-rich label-free signal profiles for different GPCR pathways, providing a more accurate pathway separation. For this, we measured real-time full-angular SPR response curves for Gs, Gq, and Gi signaling pathways in living cells. By selecting two of the most prominent label-free parameters: the full SPR curve angular and intensity shifts, we present how this analysis approach can separate each of the three signaling pathways in a straightforward single-step analysis setup, without concurrent use of signal inhibitors or other response modulating compounds.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cricetulus
  • Humans
  • Receptors, G-Protein-Coupled / metabolism*
  • Signal Transduction*
  • Surface Plasmon Resonance / methods*

Substances

  • Receptors, G-Protein-Coupled