Deconvolution of complex G protein-coupled receptor signaling in live cells using dynamic mass redistribution measurements.
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.
Schröder
Ralf
R
Molecular, Cellular, and Pharmacobiology Section, Institute of Pharmaceutical Biology, University of Bonn, Bonn, Germany.
Janssen
Nicole
N
Schmidt
Johannes
J
Kebig
Anna
A
Merten
Nicole
N
Hennen
Stephanie
S
Müller
Anke
A
Blättermann
Stefanie
S
Mohr-Andrä
Marion
M
Zahn
Sabine
S
Wenzel
Jörg
J
Smith
Nicola J
NJ
Gomeza
Jesús
J
Drewke
Christel
C
Milligan
Graeme
G
Mohr
Klaus
K
Kostenis
Evi
E
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D016466:CHO Cells; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D004789:Enzyme Activation; D044365:GTP-Binding Protein alpha Subunits, G12-G13 / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism; D009928:Organ Specificity; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D015398:Signal Transduction
D043562:Receptors, G-Protein-Coupled; D044365:GTP-Binding Protein alpha Subunits, G12-G13; D000262:Adenylyl Cyclases