Plasma membrane dynamics and tetrameric organisation of ABCG2 transporters in mammalian cells revealed by single particle imaging techniques

ABCG2 is one of three human ATP binding cassette (ABC) transporters involved in the export from cells of a chemically and structurally diverse range of compounds. This multidrug efflux capability, together with a broad tissue distribution in the body, means that ABCG2 exerts a range of effects on no...

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Main Authors: Wong, Kelvin, Briddon, Stephen J., Holliday, Nicholas D., Kerr, Ian D.
Format: Article
Published: Elsevier 2016
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Online Access:https://eprints.nottingham.ac.uk/34239/
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author Wong, Kelvin
Briddon, Stephen J.
Holliday, Nicholas D.
Kerr, Ian D.
author_facet Wong, Kelvin
Briddon, Stephen J.
Holliday, Nicholas D.
Kerr, Ian D.
author_sort Wong, Kelvin
building Nottingham Research Data Repository
collection Online Access
description ABCG2 is one of three human ATP binding cassette (ABC) transporters involved in the export from cells of a chemically and structurally diverse range of compounds. This multidrug efflux capability, together with a broad tissue distribution in the body, means that ABCG2 exerts a range of effects on normal physiology such as kidney urate transport, as well as contributing towards the pharmacokinetic profiles of many exogenous drugs. The primary sequence of ABCG2 contains only half the number of domains required for a functioning ABC transporter and so it must oligomerise in order to function, yet its oligomeric state in intact cell membranes remains uncharacterized. We have analysed ABCG2 in living cell membranes using a combination of fluorescence correlation spectroscopy, photon counting histogram analysis, and stepwise photobleaching to demonstrate a predominantly tetrameric structure for ABCG2 in the presence or absence of transport substrates. These results provide the essential basis for exploring pharmacological manipulation of oligomeric state as a strategy to modulate ABCG2 activity in future selective therapeutics.
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spelling nottingham-342392020-05-04T17:25:08Z https://eprints.nottingham.ac.uk/34239/ Plasma membrane dynamics and tetrameric organisation of ABCG2 transporters in mammalian cells revealed by single particle imaging techniques Wong, Kelvin Briddon, Stephen J. Holliday, Nicholas D. Kerr, Ian D. ABCG2 is one of three human ATP binding cassette (ABC) transporters involved in the export from cells of a chemically and structurally diverse range of compounds. This multidrug efflux capability, together with a broad tissue distribution in the body, means that ABCG2 exerts a range of effects on normal physiology such as kidney urate transport, as well as contributing towards the pharmacokinetic profiles of many exogenous drugs. The primary sequence of ABCG2 contains only half the number of domains required for a functioning ABC transporter and so it must oligomerise in order to function, yet its oligomeric state in intact cell membranes remains uncharacterized. We have analysed ABCG2 in living cell membranes using a combination of fluorescence correlation spectroscopy, photon counting histogram analysis, and stepwise photobleaching to demonstrate a predominantly tetrameric structure for ABCG2 in the presence or absence of transport substrates. These results provide the essential basis for exploring pharmacological manipulation of oligomeric state as a strategy to modulate ABCG2 activity in future selective therapeutics. Elsevier 2016-01-01 Article PeerReviewed Wong, Kelvin, Briddon, Stephen J., Holliday, Nicholas D. and Kerr, Ian D. (2016) Plasma membrane dynamics and tetrameric organisation of ABCG2 transporters in mammalian cells revealed by single particle imaging techniques. Biochimica et Biophysica Acta - Molecular Cell Research, 1863 (1). pp. 19-29. ISSN 0006-3002 Oligomerisation Fluorescence correlation spectroscopy ABCG2 Photon counting histogram Total internal reflection fluorescence Stepwise photobleaching http://www.sciencedirect.com/science/article/pii/S0167488915003493 doi:10.1016/j.bbamcr.2015.10.002 doi:10.1016/j.bbamcr.2015.10.002
spellingShingle Oligomerisation
Fluorescence correlation spectroscopy
ABCG2
Photon counting histogram
Total internal reflection fluorescence
Stepwise photobleaching
Wong, Kelvin
Briddon, Stephen J.
Holliday, Nicholas D.
Kerr, Ian D.
Plasma membrane dynamics and tetrameric organisation of ABCG2 transporters in mammalian cells revealed by single particle imaging techniques
title Plasma membrane dynamics and tetrameric organisation of ABCG2 transporters in mammalian cells revealed by single particle imaging techniques
title_full Plasma membrane dynamics and tetrameric organisation of ABCG2 transporters in mammalian cells revealed by single particle imaging techniques
title_fullStr Plasma membrane dynamics and tetrameric organisation of ABCG2 transporters in mammalian cells revealed by single particle imaging techniques
title_full_unstemmed Plasma membrane dynamics and tetrameric organisation of ABCG2 transporters in mammalian cells revealed by single particle imaging techniques
title_short Plasma membrane dynamics and tetrameric organisation of ABCG2 transporters in mammalian cells revealed by single particle imaging techniques
title_sort plasma membrane dynamics and tetrameric organisation of abcg2 transporters in mammalian cells revealed by single particle imaging techniques
topic Oligomerisation
Fluorescence correlation spectroscopy
ABCG2
Photon counting histogram
Total internal reflection fluorescence
Stepwise photobleaching
url https://eprints.nottingham.ac.uk/34239/
https://eprints.nottingham.ac.uk/34239/
https://eprints.nottingham.ac.uk/34239/