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

作者：Teemu Suutari, Sabrina N. Rahman, Henry F. Vischer, Dick van Iperen, Arto Merivaara, Marjo Yliperttula, Rob Leurs, Jeroen Kool, Tapani Viitala · 发表于：Analytical Chemistry · 年份：2020 · DOI：10.1021/acs.analchem.0c02652 · 被引用次数：5 · 研究领域：Receptor Mechanisms and Signaling、Monoclonal and Polyclonal Antibodies Research、Advanced Biosensing Techniques and Applications

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 G s, G q, and G i 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.