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

Publication year: 2020
Authors: Temu Suutari*, Sabrina N. Rahman, Henry F. Vischer, Dick van Iperen, Arto Merivaara, Marjo Yliperttula, Rob Leurs, Jeroen Kool, Tapani Viitala*
Affiliations:

* Corresponding authors:
Teemu Suutari − Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Finland; Amsterdam Institute for Medicines, Molecules and Systems, Vrije Universiteit Amsterdam, The Netherlands
Tapani Viitala − Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Finland
Authors:
Sabrina N. Rahman − Division of Medicinal Chemistry, Amsterdam Institute for Medicines, Molecules and Systems, Faculty of Science, Vrije Universiteit Amsterdam, The Netherlands
Henry F. Vischer − Division of Medicinal Chemistry, Amsterdam Institute for Medicines, Molecules and Systems, Faculty of Science, Vrije Universiteit Amsterdam, The Netherlands
Dick van Iperen − Precision Mechanics and Engineering Beta, Vrije Universiteit Amsterdam, The Netherlands
Arto Merivaara − Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Finland
Marjo Yliperttula - Division of Pharmaceutical Biosciences, Faculty of Pharmacy, University of Helsinki, Finland
Rob Leurs − Division of Medicinal Chemistry, Amsterdam Institute for Medicines, Molecules and Systems, Vrije Universiteit Amsterdam, The Netherlands
Jeroen Kool − Division of BioAnalytical Chemistry, Amsterdam Institute for Medicines, Molecules and Systems, Vrije Universiteit Amsterdam, The Netherlands; Centre for Analytical Sciences Amsterdam (CASA), Amsterdam, The Netherlands

Published in: Analytical Chemistry, 2020, Vol. 92, 21, p. 14509–14516
doi: 10.1021/acs.analchem.0c02652

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.


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