Imperial College London
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DrBenJones

Faculty of Medicine, Department of Metabolism, Digestion and Reproduction

Clinical Senior Lecturer in Metabolic Medicine
 
 
 
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Contact

 

ben.jones

 
 
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Location

 

Hammersmith HospitalHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Poc:2020:10.1039/d0sc02794d,
author = {Poc, P and Gutzeit, VA and Ast, J and Lee, J and Jones, BJ and D'Este, E and Mathes, B and Lehmann, M and Hodson, DJ and Levitz, J and Broichhagen, J},
doi = {10.1039/d0sc02794d},
journal = {Chemical Science},
pages = {7871--7883},
title = {Interrogating surfaceversusintracellular transmembrane receptor populations using cell-impermeable SNAP-tag substrates},
url = {http://dx.doi.org/10.1039/d0sc02794d},
volume = {11},
year = {2020}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - Employing self-labelling protein tags for the attachment of fluorescent dyes has become a routine and powerful technique in optical microscopy to visualize and track fused proteins. However, membrane permeability of the dyes and the associated background signals can interfere with the analysis of extracellular labelling sites. Here we describe a novel approach to improve extracellular labelling by functionalizing the SNAP-tag substrate benzyl guanine (“BG”) with a charged sulfonate (“SBG”). This chemical manipulation can be applied to any SNAP-tag substrate, improves solubility, reduces non-specific staining and renders the bioconjugation handle impermeable while leaving its cargo untouched. We report SBG-conjugated fluorophores across the visible spectrum, which cleanly label SNAP-fused proteins in the plasma membrane of living cells. We demonstrate the utility of SBG-conjugated fluorophores to interrogate class A, B and C G protein-coupled receptors (GPCRs) using a range of imaging approaches including nanoscopic superresolution imaging, analysis of GPCR trafficking from intra- and extracellular pools, in vivo labelling in mouse brain and analysis of receptor stoichiometry using single molecule pull down.
AU  - Poc,P
AU  - Gutzeit,VA
AU  - Ast,J
AU  - Lee,J
AU  - Jones,BJ
AU  - D'Este,E
AU  - Mathes,B
AU  - Lehmann,M
AU  - Hodson,DJ
AU  - Levitz,J
AU  - Broichhagen,J
DO  - 10.1039/d0sc02794d
EP  - 7883
PY  - 2020///
SN  - 2041-6520
SP  - 7871
TI  - Interrogating surfaceversusintracellular transmembrane receptor populations using cell-impermeable SNAP-tag substrates
T2  - Chemical Science
UR  - http://dx.doi.org/10.1039/d0sc02794d
UR  - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000555801800012&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR  - http://hdl.handle.net/10044/1/82610
VL  - 11
ER  -
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