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Protein-Protein Interactions pp 93–111Cite as

Measuring Rhodopsin-G-Protein Interactions by Surface Plasmon Resonance

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 261))

Abstract

G-protein-coupled receptors (GPCRs) initiate a variety of cellular responses to diverse array of extracellular stimuli. Surface plasmon resonance detection offers a powerful approach to the study of protein-protein interactions in real time. In this chapter we outline procedures for the immobilization of the prototype GPCR structure, rhodopsin or the G-protein α and βγ subunits, for analysis of the molecular interactions initiating G-protein signaling. The attachment of rhodopsin via its extracellular carbohydrate residues provides a convenient, and universally applicable, procedure for GPCR immobilization in a form that retains full biochemical activity and ability to interact with intracellular signaling components. SPR detection then allows for the analysis of the kinetic and equilibrium binding properties of the immobilized receptor with G-protein subunits and potentially other interacting molecules.

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Author information

Authors and Affiliations

  1. Laboratory of Cellular Biology, NIDCD, National Institutes of Health, Rockville, MD

    John Northup

Authors
  1. John Northup
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Editor information

Editors and Affiliations

  1. Department of Pharmacology, Emory University School of Medicine, Atlanta, GA

    Haian Fu

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© 2004 Humana Press Inc., Totowa, NJ

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Northup, J. (2004). Measuring Rhodopsin-G-Protein Interactions by Surface Plasmon Resonance. In: Fu, H. (eds) Protein-Protein Interactions. Methods in Molecular Biology, vol 261. Humana Press. https://doi.org/10.1385/1-59259-762-9:093

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  • DOI: https://doi.org/10.1385/1-59259-762-9:093

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-120-2

  • Online ISBN: 978-1-59259-762-8

  • eBook Packages: Springer Protocols

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