Modular transient nanoclustering of activated β2-adrenergic receptors revealed by single-molecule tracking of conformation-specific nanobodies.

Proceedings of the National Academy of Sciences of the United States of America
Rachel S GormalFrédéric A Meunier

Abstract

None of the current superresolution microscopy techniques can reliably image the changes in endogenous protein nanoclustering dynamics associated with specific conformations in live cells. Single-domain nanobodies have been invaluable tools to isolate defined conformational states of proteins, and we reasoned that expressing these nanobodies coupled to single-molecule imaging-amenable tags could allow superresolution analysis of endogenous proteins in discrete conformational states. Here, we used anti-GFP nanobodies tagged with photoconvertible mEos expressed as intrabodies, as a proof-of-concept to perform single-particle tracking on a range of GFP proteins expressed in live cells, neurons, and small organisms. We next expressed highly specialized nanobodies that target conformation-specific endogenous β2-adrenoreceptor (β2-AR) in neurosecretory cells, unveiling real-time mobility behaviors of activated and inactivated endogenous conformers during agonist treatment in living cells. We showed that activated β2-AR (Nb80) is highly immobile and organized in nanoclusters. The Gαs-GPCR complex detected with Nb37 displayed higher mobility with surprisingly similar nanoclustering dynamics to that of Nb80. Activated conformers are hig...Continue Reading

References

Jun 3, 1993·Nature·C Hamers-CastermanR Hamers
Jan 12, 2002·The American Journal of Pathology·Carlo MinettiGiuseppe Cordone
Mar 7, 2002·Developmental Cell·William A MohlerBenjamin Podbilewicz
Aug 28, 2003·Médecine sciences : M/S·Christelle Gally, Jean-Louis Bessereau
Jul 1, 2004·Proceedings of the National Academy of Sciences of the United States of America·Antje KepplerKai Johnsson
Dec 30, 2004·The Journal of Biological Chemistry·Wakako ShimoiKatsuko Tani
Aug 10, 2006·Nature Methods·Michael J RustXiaowei Zhuang
Aug 12, 2006·Science·Eric BetzigHarald F Hess
Sep 19, 2006·Biophysical Journal·Samuel T HessMichael D Mason
Oct 14, 2006·Annual Review of Physiology·Catherine A C MooreJeffrey L Benovic
Oct 25, 2006·Nature Methods·Ulrich RothbauerHeinrich Leonhardt
Dec 1, 2007·WormBook : the Online Review of C. Elegans Biology·Benjamin Podbilewicz
Jan 15, 2008·Nature Methods·Suliana ManleyJennifer Lippincott-Schwartz
May 29, 2008·Journal of Cell Science·Matthew KirkhamRobert G Parton
Jun 7, 2008·ACS Chemical Biology·Georgyi V LosKeith V Wood
May 22, 2009·Nature·Daniel M RosenbaumBrian K Kobilka
May 28, 2009·Proceedings of the National Academy of Sciences of the United States of America·Xiao Jie YaoBrian Kobilka
Mar 6, 2010·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Anindya Ghosh-RoyAndrew D Chisholm
May 8, 2010·Science·Meital Oren-SuissaBenjamin Podbilewicz
Jul 20, 2010·Toxicon : Official Journal of the International Society on Toxinology·Jacqueline M TremblayCharles B Shoemaker
Aug 18, 2010·Biophysical Journal·Gregory GiannoneLaurent Cognet
Oct 15, 2010·Protein Science : a Publication of the Protein Society·Marta H KubalaBrett M Collins
Jan 14, 2011·Nature·Søren G F RasmussenBrian K Kobilka
Apr 20, 2011·Biochimica Et Biophysica Acta·Sourav GangulyAmitabha Chattopadhyay
Jun 1, 2011·Journal of Cell Science·Emmanuel BoucrotRobert G Parton
Jun 15, 2011·Nature Chemical Biology·Akash GulyaniKlaus M Hahn
Jul 21, 2011·Nature·Søren G F RasmussenBrian K Kobilka
Aug 11, 2011·PloS One·Anette I ÖbergTore Bengtsson
Aug 13, 2011·The Journal of Biological Chemistry·Callista B HarperFrederic A Meunier
Oct 23, 2012·Methods in Molecular Biology·Grégory GiannoneLaurent Cognet
Nov 21, 2012·Trends in Cell Biology·Callista B HarperFrédéric A Meunier
Jan 24, 2013·Nature Reviews. Molecular Cell Biology·Robert G Parton, Miguel A del Pozo
Feb 5, 2013·Cell·Rie NygaardBrian K Kobilka
Mar 19, 2013·Annual Review of Biochemistry·Serge Muyldermans
Mar 22, 2013·Nature·Roshanak IrannejadMark von Zastrow
Jul 6, 2013·Science·Ibrahim I CisseXavier Darzacq

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Citations

Dec 8, 2020·The FEBS Journal·Alexey Bondar, Josef Lazar
Jun 20, 2021·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Xiaorong PanTong Wang
Jul 14, 2021·Chembiochem : a European Journal of Chemical Biology·Hideaki Yoshimura
Oct 12, 2021·The Journal of Cell Biology·Harriet P LoRobert G Parton

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