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Single-molecule resolution of G protein-coupled receptor (GPCR) complexes.

Jonas, KC; Huhtaniemi, I; Hanyaloglu, AC (2016) Single-molecule resolution of G protein-coupled receptor (GPCR) complexes. Methods in Cell Biology, 132. pp. 55-72. ISSN 0091-679X https://doi.org/10.1016/bs.mcb.2015.11.005
SGUL Authors: Jonas, Kim Carol

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Abstract

The organization of G protein-coupled receptors (GPCRs) into dimers and higher-order oligomers has provided a potential mechanistic system in defining complex GPCR responses. Despite being studied for nearly 20years it has, and still is, been an area of controversy. Although technology has developed to quantitatively measure these associations in real time, identify the structural interfaces and even systems to understand the physiological significance of di/oligomerization, key questions remain outstanding including the role of each individual complex from the monomer to the higher-order oligomer, in their native system. Recently, single-molecule microscopy approaches have provided the tools to directly visualize individual GPCRs in dimers and oligomers, though as with any technological development each have their advantages and limitations. This chapter will describe these recent developments in single-molecule fluorescent microscopy, focusing on our recent application of super-resolution imaging of the GPCR for the luteinizing hormone/chorionic gonadotropin to quantify GPCR monomers and formation of protomers in to dimers and distinct oligomeric forms. We present our approach, considerations, strategy, and challenges to visualize this receptor beyond the light diffraction limit via photoactivated localization microscopy with photoactivatable dyes. The addition of super-resolution approaches to the GPCR "nano-tool kit" will pave the way for novel avenues to answer outstanding questions regarding the existence and significance of these complexes to GPCR signaling.

Item Type: Article
Additional Information: © 2016 Elsevier Inc. All rights reserved.
Keywords: Dimerization, G protein-coupled receptors, Oligomerization, Signaling, Single molecule, Super-resolution imaging, Developmental Biology, 0601 Biochemistry And Cell Biology
SGUL Research Institute / Research Centre: Academic Structure > Institute of Medical & Biomedical Education (IMBE)
Academic Structure > Institute of Medical & Biomedical Education (IMBE) > Centre for Biomedical Education (INMEBE)
Journal or Publication Title: Methods in Cell Biology
ISSN: 0091-679X
Language: eng
Dates:
DateEvent
2016Published
Publisher License: Publisher's own licence
PubMed ID: 26928539
Go to PubMed abstract
URI: https://openaccess.sgul.ac.uk/id/eprint/107823
Publisher's version: https://doi.org/10.1016/bs.mcb.2015.11.005

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