Deconvolution of complex G protein-coupled receptor signaling in live cells using dynamic mass redistribution measurements

• Published in: Nature biotechnology Vol. 28; no. 9; pp. 943 - 949
• Main Authors: Mohr, Klaus, Kostenis, Evi, Schröder, Ralf, Janssen, Nicole, Schmidt, Johannes, Kebig, Anna, Merten, Nicole, Hennen, Stephanie, Müller, Anke, Blättermann, Stefanie, Mohr-Andrä, Marion, Zahn, Sabine, Wenzel, Jörg, Smith, Nicola J, Gomeza, Jesús, Drewke, Christel, Milligan, Graeme
• Format: Journal Article
• Language: English
• Published: New York, NY Nature Publishing Group 01.09.2010
• Subjects: Adenylyl Cyclases - metabolism; Animals; Biological and medical sciences; Biosensing Techniques - methods; Biosensors; Biotechnology; Cell Survival; Cellular biology; Cellular proteins; CHO Cells; Cricetinae; Cricetulus; Diverse techniques; Enzyme Activation; Fundamental and applied biological sciences. Psychology; G proteins; GTP-Binding Protein alpha Subunits, G12-G13 - metabolism; HEK293 Cells; Humans; Keratinocytes - metabolism; Molecular and cellular biology; Observations; Organ Specificity; Properties; Proteins; Receptors, G-Protein-Coupled - metabolism; Signal Transduction; United Kingdom; Germany; Analysis method; Signaling pathway; G protein coupled receptor; Biosensor; Cell signaling
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/nbt.1671

Label-free biosensor technology based on dynamic mass redistribution (DMR) of cellular constituents promises to translate GPCR signaling into complex optical 'fingerprints' in real time in living cells. Here we present a strategy to map cellular mechanisms that define label-free responses, and we compare DMR technology with traditional second-messenger assays that are currently the state of the art in GPCR drug discovery. The holistic nature of DMR measurements enabled us to (i) probe GPCR functionality along all four G-protein signaling pathways, something presently beyond reach of most other assay platforms; (ii) dissect complex GPCR signaling patterns even in primary human cells with unprecedented accuracy; (iii) define heterotrimeric G proteins as triggers for the complex optical fingerprints; and (iv) disclose previously undetected features of GPCR behavior. Our results suggest that DMR technology will have a substantial impact on systems biology and systems pharmacology as well as for the discovery of drugs with novel mechanisms.