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

被引：207

作者：

Schroeder, Ralf ^{[1
]}

Janssen, Nicole ^{[2
]}

Schmidt, Johannes ^{[1
]}

Kebig, Anna ^{[2
]}

Merten, Nicole ^{[1
]}

Hennen, Stephanie ^{[1
]}

Mueller, Anke ^{[1
]}

Blaettermann, Stefanie ^{[1
]}

Mohr-Andrae, Marion ^{[2
]}

Zahn, Sabine ^{[3
]}

Wenzel, Joerg ^{[3
]}

Smith, Nicola J. ^{[4
]}

Gomeza, Jesus ^{[1
]}

Drewke, Christel ^{[1
]}

Milligan, Graeme ^{[4
]}

Mohr, Klaus ^{[2
]}

Kostenis, Evi ^{[1
]}

机构：

[1] Univ Bonn, Inst Pharmazeut Biol, Mol Cellular & Pharmacobiol Sect, D-5300 Bonn, Germany

[2] Univ Bonn, Pharmacol & Toxicol Sect, Inst Pharm, D-5300 Bonn, Germany

[3] Univ Bonn, Dept Dermatol, D-5300 Bonn, Germany

[4] Univ Glasgow, Mol Pharmacol Grp, Fac Biomed & Life Sci, Glasgow, Lanark, Scotland

来源：

NATURE BIOTECHNOLOGY | 2010年 / 28卷 / 09期

关键词：

DRUG DISCOVERY; 7-TRANSMEMBRANE RECEPTORS; OPTICAL BIOSENSOR; FATTY-ACIDS; ACTIVATION; LIGANDS; AGONIST; GPR55; ASSAY; PHARMACOLOGY;

D O I：

10.1038/nbt.1671

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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. (C) 2010 Nature America, Inc. All rights reserved.

引用

页码：943 / 950

页数：8

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