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

共 50 条

[1] Dualsteric GPCR targeting: a novel route to binding and signaling pathway selectivity [J].

Antony, Johannes ;

Kellershohn, Kerstin ;

Mohr-Andrae, Marion ;

Kebig, Anna ;

Prilla, Stefanie ;

Muth, Mathias ;

Heller, Eberhard ;

Disingrini, Teresa ;

Dallanoce, Clelia ;

Bertoni, Simona ;

Schrobang, Jasmin ;

Traenkle, Christian ;

Kostenis, Evi ;

Christopoulos, Arthur ;

Hoeltje, Hans-Dieter ;

Barocelli, Elisabetta ;

De Amici, Marco ;

Holzgrabe, Ulrike ;

Mohr, Klaus .

FASEB JOURNAL, 2009, 23 (02) :442-450

[2] β-Arrestin-mediated activation of MAPK by inverse agonists reveals distinct active conformations for G protein-coupled receptors [J].

Azzi, M ;

Charest, PG ;

Angers, S ;

Rousseau, G ;

Kohout, T ;

Bouvier, M ;

Piñeyro, G .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (20) :11406-11411

[3] Multiple GPCR conformations and signalling pathways: implications for antagonist affinity estimates [J].

Baker, Jillian G. ;

Hill, Stephen J. .

TRENDS IN PHARMACOLOGICAL SCIENCES, 2007, 28 (08) :374-381

[4] The genetic design of signaling cascades to record receptor activation [J].

Barnea, Gilad ;

Strapps, Walter ;

Herrada, Gilles ;

Berman, Yemiliya ;

Ong, Jane ;

Kloss, Brian ;

Axel, Richard ;

Lee, Kevin J. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (01) :64-69

[5] Versatility of GPCR recognition by drugs: from biological implications to therapeutic relevance [J].

Bosier, Barbara ;

Hermans, Emmanuel .

TRENDS IN PHARMACOLOGICAL SCIENCES, 2007, 28 (08) :438-446

[6] The orphan G protein-coupled receptor GPR40 is activated by medium and long chain fatty acids [J].

Briscoe, CP ;

Tadayyon, M ;

Andrews, JL ;

Benson, WG ;

Chambers, JK ;

Eilert, MM ;

Ellis, C ;

Elshourbagy, NA ;

Goetz, AS ;

Minnick, DT ;

Murdock, PR ;

Sauls, HR ;

Shabon, U ;

Spinage, LD ;

Strum, JC ;

Szekeres, PG ;

Tan, KB ;

Way, JM ;

Ignar, DM ;

Wilson, S ;

Muir, AI .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (13) :11303-11311

[7]

BURFORD NT, 1995, J PHARMACOL EXP THER, V274, P134

[8] Discovery of Potent and Selective Agonists for the Free Fatty Acid Receptor 1 (FFA1/GPR40), a Potential Target for the Treatment of Type II Diabetes [J].

Christiansen, Elisabeth ;

Urban, Christian ;

Merten, Nicole ;

Liebscher, Kathrin ;

Karlsen, Kasper K. ;

Hamacher, Alexandra ;

Spinrath, Andreas ;

Bond, Andrew D. ;

Drewke, Christel ;

Ullrich, Susanne ;

Kassack, Matthias U. ;

Kostenis, Evi ;

Ulven, Trond .

JOURNAL OF MEDICINAL CHEMISTRY, 2008, 51 (22) :7061-7064

[9] A 100K well screen for a muscarinic receptor using the Epic® label-free system - a reflection on the benefits of the label-free approach to screening seven-transmembrane receptors [J].

Dodgson, K. ;

Gedge, L. ;

Murray, D. C. ;

Coldwell, M. .

JOURNAL OF RECEPTORS AND SIGNAL TRANSDUCTION, 2009, 29 (3-4) :163-172

[10] Functional G protein-coupled receptor assays for primary and secondary screening [J].

Eglen, RM .

COMBINATORIAL CHEMISTRY & HIGH THROUGHPUT SCREENING, 2005, 8 (04) :311-318

← 1 2 3 4 5 →