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Methods to Immobilize GPCR on the Surface of SPR Sensors

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Book cover G Protein-Coupled Receptor Screening Assays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1272))

Abstract

The G protein-coupled receptors (GPCRs) form one of the largest membrane receptor families. The nature of the ligands that interact with these receptors is highly diverse; they include light, peptides and hormones, neurotransmitters, and small molecular weight compounds. The GPCRs are involved in a wide variety of physiological processes and thus hold considerable therapeutic potential.

GPCR function is usually determined in cell-based assays, whose complexity nonetheless limits their use. The use of alternative, cell-free assays is hampered by the difficulties in purifying these seven-transmembrane domain receptors without altering their functional properties. Several methods have been proposed to immobilize GPCR on biosensor surfaces which use antibodies or avidin-/biotin-based capture procedures, alone or with reconstitution of the GPCR physiological microenvironment. Here we propose a method for GPCR immobilization in their native membrane microenvironment that requires no manipulation of the target receptor and maintains the many conformations GPCR can adopt in the cell membrane.

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Acknowledgments

We thank the people at the Chemokine SIgnaling Group for much of the work that contributed to this review, especially Beatriz Vega, Pilar Lucas, and Jorge Villaverde, and to C. Bastos and C. Mark for the secretarial and editorial assistance, respectively. This work was supported in part by grants from the Spanish Ministry of Science and Innovation (SAF 2011-27370), the RETICS Program (RD12/0009/009; RIER), and the Madrid regional government (S2010/BMD-2350; RAPHYME).

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Authors and Affiliations

  1. Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB/CSIC), Darwin 3, Campus de Cantoblanco, Madrid, 28049, Spain

    Laura Martínez-Muñoz, Rubén Barroso, Anabel Guedán Paredes, Mario Mellado & José Miguel Rodríguez-Frade

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  1. Laura Martínez-Muñoz
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  2. Rubén Barroso
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  3. Anabel Guedán Paredes
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  4. Mario Mellado
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  5. José Miguel Rodríguez-Frade
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Corresponding author

Correspondence to José Miguel Rodríguez-Frade .

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Editors and Affiliations

  1. IBB – Institute for Biotechnology and Bioengineering, Lisbon, Portugal

    Duarte Miguel F. Prazeres  & Sofia Aires M. Martins  & 

  2. Department of Bioengineering, Instituto Superior Técnico Universidade de Lisboa, Lisbon, Portugal

    Duarte Miguel F. Prazeres

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Martínez-Muñoz, L., Barroso, R., Paredes, A.G., Mellado, M., Rodríguez-Frade, J.M. (2015). Methods to Immobilize GPCR on the Surface of SPR Sensors. In: Prazeres, D.M.F., Martins, S.A.M. (eds) G Protein-Coupled Receptor Screening Assays. Methods in Molecular Biology, vol 1272. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-2336-6_12

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  • DOI: https://doi.org/10.1007/978-1-4939-2336-6_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-2335-9

  • Online ISBN: 978-1-4939-2336-6

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