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Super-resolution imaging as a method to Study GPCR dimers and higher-order oligomers

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Title: Super-resolution imaging as a method to Study GPCR dimers and higher-order oligomers
Authors: Jonas, KC
Hanyaloglu, AC
Item Type: Chapter
Abstract: The study of G protein-coupled receptor (GPCR) dimers and higher order oligomers has unveiled mechanisms for receptors to diversify signaling and potentially uncover novel therapeutic targets. The functional and clinical significance of these receptorreceptor associations has been facilitated by the development of techniques and protocols, enabling researchers to unpick their function from the molecular interfaces, to demonstrating functional significance in vivo, in both health and disease. Here we describe our methodology to study GPCR oligomerization at the single molecule level via super-resolution imaging. Specifically, we have employed photoactivated localization microscopy, with photoactivatable dyes (PD-PALM) to visualize the spatial organization of these complexes to <10nm resolution, and the quantitation of GPCR monomer, dimer and oligomer in both homomeric and heteromeric forms. We provide guidelines on optimal sample preparation, imaging parameters and necessary controls for resolving and quantifying single molecule data. Finally, we discuss advantages and limitations of this imaging technique and its potential future applications to the study of GPCR function.
Issue Date: 2-Aug-2018
URI: http://hdl.handle.net/10044/1/71356
DOI: https://doi.org/10.1007/978-1-4939-8576-0_21
Publisher: Springer
Start Page: 329
End Page: 343
Journal / Book Title: Receptor-receptor interactions in the central nervous system
Copyright Statement: © Springer Science+Business Media, LLC, part of Springer Nature 2018
Sponsor/Funder: Biotechnology and Biological Sciences Research Council (BBSRC)
Funder's Grant Number: BB/I008004/1
Article Number: 21
Online Publication Date: 2018-08-02
Appears in Collections:Department of Surgery and Cancer