Abstract
Signal transduction by G-protein coupled receptors (GPCRs) underpins a multitude of physiological processes. Ligand recognition by these receptors leads to activation of a generic molecular switch involving heterotrimeric G-proteins and guanine nucleotides. With growing interest and commercial investment in GPCRs in areas such as drug targets, orphan receptors, high-throughput screening of drugs, biosensors etc., greater attention will focus on assay development to allow for miniaturization, ultrahigh throughput, and eventually, microarray/biochip assay formats that will require nanotechnology-based approaches. Stable, robust, cell-free signaling assemblies comprising receptor and appropriate molecular switching components will form the basis of future GPCR/G-protein platforms which should be adaptable for such applications as microarrays and biosensors. This chapter focuses on cell-free GPCR assay nanotechnologies and describes some molecular biological approaches for the construction of more sophisticated, surface-immobilized, homogeneous, functional GPCR sensors. The latter points should greatly extend the range of applications to which technologies based on GPCRs could be applied.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Abbreviations
- AFM:
-
atomic force microscope
- AFM:
-
atomic force microscopy
- ATP:
-
adenosine triphosphate
- FCS:
-
fluorescence correlation spectroscopy
- FP:
-
fluorescence polarization
- FPR:
-
N-formyl peptide receptor
- GDP:
-
guanosine diphosphate
- GFP:
-
green fluorescent protein
- GPCR:
-
G-protein coupled receptor
- GTP:
-
guanosine triphosphate
- HTS:
-
high throughput screening
- IMAC:
-
immobilized metal ion affinity chromatography
- NTA:
-
nitrilotriacetate
- PLC:
-
phospholipase C
- PWR:
-
plasmon-waveguide resonance
- SPR:
-
surface plasmon resonance
- TCM:
-
tetracysteine motif
- TIRF:
-
total internal reflection fluorescence
- TMR:
-
tetramethylrhodamine
- TPE-FCCS:
-
two-photon excitation fluorescence cross-correlation spectroscopy
- YFP:
-
yellow fluorescent protein
- cAMP:
-
cyclic adenosine monophosphate
References
A. Wise, K. Gearing, S. Rees: Target validation of G-protein coupled receptors, Drug Discov. Today 7, 235–246 (2002)
S.G. Rasmussen, H.J. Choi, D.M. Rosenbaum, T.S. Kobilka, F.S. Thian, P.C. Edwards: Crystal structure of the human beta(2) adrenergic G-protein-coupled receptor, Nature 450, 383–387 (2007)
K.L. Pierce, R.T. Premont, R.J. Lefkowitz: Seven-transmembrane receptors, Nat. Rev. Mol. Cell Biol. 3, 639–650 (2002)
D.K. Vassilatis, J.G. Hohmann, H. Zeng, F. Li, J.E. Ranchalis, M.T. Mortrud: The G protein-coupled receptor repertoires of human and mouse, Proc. Natl. Acad. Sci. USA 100, 4903–4908 (2003)
V.V. Verkhusha, K.A. Lukyanov: The molecular properties and applications of anthozoa fluorescent proteins and chromoproteins, Nat. Biotechnol. 22, 289–296 (2004)
T.H. Cooper, W.R. Leifert, R.V. Glatz, E.J. McMurchie: Expression and characterisation of functional lanthanide binding tags fused to a Gα-protein and muscarinic (M2) receptor, J. Bionanosci. 2(1), 27–34 (2008)
S.R. Adams, R.E. Campbell, L.A. Gross, B.R. Martin, G.K. Walkup, Y. Yao: New biarsenical ligands and tetracysteine motifs for protein labeling in vitro and in vivo: Synthesis and biological applications, J. Am. Chem. Soc. 124, 6063–6076 (2002)
C. Hoffmann, G. Gaietta, M. Bunemann, S.R. Adams, S. Oberdorff-Maass, B. Behr: A FlAsH-based FRET approach to determine G protein-coupled receptor activation in living cells, Nat. Methods 2, 171–176 (2005)
M.P. Caulfield, N.J. Birdsall: International union of pharmacology. XVII. classification of muscarinic acetylcholine receptors, Pharmacol. Rev. 50, 279–290 (1998)
A.M. Liu, M.K. Ho, C.S. Wong, J.H. Chan, A.H. Pau, Y.H. Wong: Gα(16/z) chimeras efficiently link a wide range of G protein-coupled receptors to calcium mobilization, J. Biomol. Screen. 8, 39–49 (2003)
A. Hazari, V. Lowes, J.H. Chan, C.S. Wong, M.K. Ho, Y.H. Wong: Replacement of the α 5 helix of Gα 16 with gα s-specific sequences enhances promiscuity of Gα 16 toward Gs-coupled receptors, Cell Signal. 16, 51–62 (2004)
T.T. Amatruda III, D.A. Steele, V.Z. Slepak, M.I. Simon: Gα 16, a G protein α subunit specifically expressed in hematopoietic cells, Proc. Natl. Acad. Sci. USA 88, 5587–5591 (1991)
S. Offermanns, M.I. Simon: Gα 15 and Gα 16 couple a wide variety of receptors to phospholipase C, J. Biol. Chem. 270, 15175–15180 (1995)
X. Zhu, L. Birnbaumer: G protein subunits and the stimulation of phospholipase C by Gs- and Gi-coupled receptors: Lack of receptor selectivity of Gα(16) and evidence for a synergic interaction between Gβγ and the α subunit of a receptor activated G protein, Proc. Natl. Acad. Sci. USA 93, 2827–2831 (1996)
G. Milligan, F. Marshall, S. Rees: G16 as a universal G protein adapter: Implications for agonist screening strategies, Trends Pharmacol. Sci. 17, 235–237 (1996)
J.W. Lee, S. Joshi, J.S. Chan, Y.H. Wong: Differential coupling of μ-, δ-, and κ-opioid receptors to Gα 16-mediated stimulation of phospholipase C, J. Neurochem. 70, 2203–2211 (1998)
E. Kostenis: Is Gα 16 the optimal tool for fishing ligands of orphan G-protein-coupled receptors?, Trends Pharmacol. Sci. 22, 560–564 (2001)
B.R. Conklin, Z. Farfel, K.D. Lustig, D. Julius, H.R. Bourne: Substitution of three amino acids switches receptor specificity of Gq α to that of Gi α, Nature 363, 274–276 (1993)
B.R. Conklin, P. Herzmark, S. Ishida, T.A. Voyno-Yasenetskaya, Y. Sun, Z. Farfel: Carboxyl-terminal mutations of Gq α and Gs α that alter the fidelity of receptor activation, Mol. Pharmacol. 50, 885–890 (1996)
E. Kostenis, F.Y. Zeng, J. Wess: Functional characterization of a series of mutant G protein α q subunits displaying promiscuous receptor coupling properties, J. Biol. Chem. 273, 17886–17892 (1998)
S.M. Mody, M.K. Ho, S.A. Joshi, Y.H. Wong: Incorporation of Gα(z)-specific sequence at the carboxyl terminus increases the promiscuity of Gα(16) toward G(i)-coupled receptors, Mol. Pharmacol. 57, 13–23 (2000)
C.H. Klaassen, P.H. Bovee-Geurts, G.L. Decaluwe, W.J. DeGrip: Large-scale production and purification of functional recombinant bovine rhodopsin with the use of the baculovirus expression system, Biochem. J. 342, 293–300 (1999)
D. Ott, Y. Neldner, R. Cebe, I. Dodevski, A. Pluckthun: Engineering and functional immobilization of opioid receptors, Protein Eng. Design Sel. 18, 153–160 (2005)
R.V. Glatz, W.R. Leifert, T.H. Cooper, K. Bailey, C.S. Barton, A.S. Martin: Molecular engineering of G protein-coupled receptors and G proteins for cell-free biosensing, Aust. J. Chem. 60, 309 (2007)
C.S. Barton, R.V. Glatz, A.S. Martin, L. Waniganayake, E.J. McMurchie, W.R. Leifert: Interaction of self-assembled monolayers incorporating NTA disulfide with multilength histidine-tagged Gα i1 subunits, J. Bionanosci. 1, 22–30 (2007)
P. Stenlund, G.J. Babcock, J. Sodroski, D.G. Myszka: Capture and reconstitution of G protein-coupled receptors on a biosensor surface, Anal. Biochem. 316, 243–250 (2003)
J.C. Owicki: Fluorescence polarization and anisotropy in high throughput screening: Perspectives and primer, J. Biomol. Screen. 5, 297–306 (2000)
C.J. Daly, J.C. McGrath: Fluorescent ligands, antibodies, and proteins for the study of receptors, Pharmacol. Ther. 100, 101–118 (2003)
P. Banks, M. Harvey: Considerations for using fluorescence polarization in the screening of G protein-coupled receptors, J. Biomol. Screen. 7, 111–117 (2002)
J.L. Swift, M.C. Burger, D. Massotte, T.E. Dahms, D.T. Cramb: Two-photon excitation fluorescence cross-correlation assay for ligand-receptor binding: Cell membrane nanopatches containing the human micro-opioid receptor, Anal. Chem. 79, 6783–6791 (2007)
A. Waller, P. Simons, E.R. Prossnitz, B.S. Edwards, L.A. Sklar: High throughput screening of G-protein coupled receptors via flow cytometry, Comb. Chem. High Throughput Screen. 6, 389–397 (2003)
T. Mirzabekov, H. Kontos, M. Farzan, W. Marasco, J. Sodroski: Paramagnetic proteoliposomes containing a pure, native, and oriented seven-transmembrane segment protein, CCR5, Nat. Biotechnol. 18, 649–654 (2000)
K.L. Martinez, B.H. Meyer, R. Hovius, K. Lundstrom, H. Vogel: Ligand binding to G protein-coupled receptors in tethered cell membranes, Langmuir 19, 10925–10929 (2003)
Y. Fang, A.G. Frutos, J. Lahiri: Membrane protein microarrays, J. Am. Chem. Soc. 124, 2394–2395 (2002)
Y. Fang, A.G. Frutos, B. Webb, Y. Hong, A. Ferrie, F. Lai: Membrane biochips, BioTechniques Supplement, 62–65 (2002)
Y. Fang, A.G. Frutos, J. Lahiri: G-protein-coupled receptor microarrays, ChemBioChem 3, 987–991 (2002)
Y. Fang, J. Peng, A.M. Ferrie, R.S. Burkhalter: Air-stable G protein-coupled receptor microarrays and ligand binding characteristics, Anal. Chem. 78, 149–155 (2006)
N.M. Rao, V. Silin, K.D. Ridge, J.T. Woodward, A.L. Plant: Cell membrane hybrid bilayers containing the G-protein-coupled receptor CCR5, Anal. Biochem. 307, 117–130 (2002)
O. Karlsson, L. Stefan: Flow-mediated on-surface reconsititution of G-protein coupled receptors for applications in surface plasmon resonance biosensors, Anal. Biochem. 300, 132–138 (2002)
G. Tollin, Z. Salamon, V.J. Hruby: Techniques: Plasmon-waveguide resonance (PWR) spectroscopy as a tool to study ligand-GPCR interactions, Trends Pharmacol. Sci. 24, 655–659 (2003)
S. Devanathan, Z. Yao, Z. Salamon, B. Kobilka, G. Tollin: Plasmon-waveguide resonance studies of ligand binding to the human beta 2-adrenergic receptor, Biochemistry 43, 3280–3288 (2004)
I.D. Alves, S.M. Cowell, Z. Salamon, S. Devanathan, G. Tollin, V.J. Hruby: Different structural states of the proteolipid membrane are produced by ligand binding to the human delta-opioid receptor as shown by plasmon-waveguide resonance spectroscopy, Mol. Pharmacol. 65, 1248–1257 (2004)
T.Z. Wu: A piezoelectric biosensor as an olfactory receptor for odour detection: Electronic nose, Biosens. Bioelectron. 14, 9–18 (1999)
G. Milligan: Principles: Extending the utility of [35S]GTPγS binding assays, Trends Pharmacol. Sci. 24, 87–90 (2003)
C. Harrison, J.R. Traynor: The [35S]GTPγS binding assay: Approaches and applications in pharmacology, Life Sci. 74, 489–508 (2003)
M. Bunemann, M. Frank, M.J. Lohse: Gi protein activation in intact cells involves subunit rearrangement rather than dissociation, Proc. Natl. Acad. Sci. USA 100, 16077–16082 (2003)
C. Bieri, O.P. Ernst, S. Heyse, K.P. Hofmann, H. Vogel: Micropatterned immobilization of a G protein-coupled receptor and direct detection of G protein activation, Nat. Biotechnol. 17, 1105–1108 (1999)
L.A. Sklar, J. Vilven, E. Lynam, D. Neldon, T.A. Bennett, E. Prossnitz: Solubilization and display of G protein-coupled receptors on beads for real-time fluorescence and flow cytometric analysis, BioTechniques 28, 975–976 (2000)
P.C. Simons, M. Shi, T. Foutz, D.F. Cimino, J. Lewis, T. Buranda: Ligand-receptor-G-protein molecular assemblies on beads for mechanistic studies and screening by flow cytometry, Mol. Pharmacol. 64, 1227–1238 (2003)
A. Waller, P.C. Simons, S.M. Biggs, B.S. Edwards, E.R. Prossnitz, L.A. Sklar: Techniques: GPCR assembly, pharmacology and screening by flow cytometry, Trends Pharmacol. Sci. 25, 663–669 (2004)
T.A. Bennett, T.A. Key, V.V. Gurevich, R. Neubig, E.R. Prossnitz, L.A. Sklar: Real-time analysis of G protein-coupled receptor reconstitution in a solubilized system, J. Biol. Chem. 276, 22453–22460 (2001)
X. Michalet, F.F. Pinaud, L.A. Bentolila, J.M. Tsay, S. Doose, J.J. Li: Quantum dots for live cells, in vivo imaging, and diagnostics, Science 307, 538–544 (2005)
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer-Verlag
About this chapter
Cite this chapter
Leifert, W.R., Cooper, T.H., Bailey, K. (2010). G-Protein Coupled Receptors: Progress in Surface Display and Biosensor Technology. In: Bhushan, B. (eds) Springer Handbook of Nanotechnology. Springer Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-02525-9_17
Download citation
DOI: https://doi.org/10.1007/978-3-642-02525-9_17
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-02524-2
Online ISBN: 978-3-642-02525-9
eBook Packages: EngineeringEngineering (R0)