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• W2886690789 abstract "G protein-coupled receptors (GPCRs) modulate cellular signaling pathways, including differentiation, proliferation, hormonal regulation, and neuronal activity. Therefore, it is not surprising that almost 50% of the drugs available in the pharmaceutical market target GPCRs. Recently, an emerging body of evidence has proven the formation of GPCR dimers and even higher order oligomers. For neurodegenerative diseases, such as Parkinson’s or Alzheimer’s disease, it is crucial to characterize these receptor–receptor interactions in the brain to elucidate their role in neuronal disease-relevant processes. As a first step, a robust in cellulo assay is essential to identify and characterize specific GPCR–GPCR interactions. In the past 20 years, considerable efforts have been directed towards the development of GPCR dimerization screening assays to evaluate these receptor–receptor interactions in living cells. Interestingly, most of the approaches employ noninvasive fluorescence- and luminescence-based assays. Here, we present an efficient strategy to study GPCR dimerization dynamics, namely a protein complementation assay (PCA) based on the reconstitution of a luminescent protein, the NanoLuciferase (NL). Thus, GPCRs of interest are fused to complementary NL fragments which upon GPCR dimerization may reconstitute to a functional reporter, of which activity can be measured. The experimental procedure takes 2–4 days to complete, depending on the cell type and complexity of the experimental setup. In contrast to alternative protein complementation assays (also described in this book chapter), this method can also be implemented to analyze the kinetics of ligand-dependent modulation of dimerization, broadening its application potential. Additionally, high throughput screenings can also be performed, which is highly relevant given the growing interest and effort to identify small molecule drugs that can target disease-relevant dimers (or even selectively alter GPCR dimer function)." @default.
• W2886690789 created "2018-08-22" @default.
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• W2886690789 date "2018-01-01" @default.
• W2886690789 modified "2023-10-07" @default.
• W2886690789 title "Assessing GPCR Dimerization in Living Cells: Comparison of the NanoBiT Assay with Related Bioluminescence- and Fluorescence-Based Approaches" @default.
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• W2886690789 doi "https://doi.org/10.1007/978-1-4939-8576-0_15" @default.
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