TY - JOUR
T1 - Label-Free Analysis with Multiple Parameters Separates G Protein-Coupled Receptor Signaling Pathways
AU - Suutari, Teemu
AU - Rahman, Sabrina N.
AU - Vischer, Henry F.
AU - Van Iperen, Dick
AU - Merivaara, Arto
AU - Yliperttula, Marjo
AU - Leurs, Rob
AU - Kool, Jeroen
AU - Viitala, Tapani
N1 - Funding Information:
This research was funded by the Doctoral Programme in Materials Research and Nanosciences at University of Helsinki, Academy of Finland (T.V.: 294309; M.Y.: 315409 and 315406) and supported by grants from the Finnish Pharmaceutical Society (T.S.), the Finnish Cultural Foundation (T.S.) and Business Finland (M.Y. and T.V.: Dnro: 1842/31/2019). The authors would like to also thank the Division of BioAnalytical Chemistry and Medicinal Chemistry at Vrije Universiteit Amsterdam and BioPharm Enterprises Limited, U.K. for their financial support. S.N.R., H.F.V., and R.L. acknowledge the Netherlands Organisation for Scientific Research (NWO) for financial support (TOPPUNT, “7 ways to 7TMR modulation (7-to-7)”, 718.014.002). R.L. and H.F.V. participated in the European Research Network on Signal Transduction (ERNEST) [COST ACTION CA 18133]. Last, we thank Dr. Walis Jones from BioPharm Enterprises Ltd. for his support with the RWG measurements and for his comments to the paper.
Publisher Copyright:
© 2020 American Chemical Society.
PY - 2020/11/3
Y1 - 2020/11/3
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85096119040&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.0c02652
DO - 10.1021/acs.analchem.0c02652
M3 - Article
AN - SCOPUS:85096119040
SN - 0003-2700
VL - 92
SP - 14509
EP - 14516
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 21
ER -