Sulfonated red and far-red rhodamines to visualize SNAP- and Halo-tagged cell surface proteins
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
The (in)ability to permeate membranes is a key feature of chemical biology probes that defines their suitability for specific applications. Here we report sulfonated rhodamines that endow xanthene dyes with cellular impermeability for analysis of surface proteins. We fuse charged sulfonates to red and far-red dyes to obtain Sulfo549 and Sulfo646, respectively, and further link these to benzylguanine and choloralkane substrates for SNAP-tag and Halo-tag labelling. Sulfonated rhodamine-conjugated fluorophores maintain desirable photophysical properties, such as brightness and photostability. While transfected cells with a nuclear localized SNAP-tag remain unlabelled, extracellular exposed tags can be cleanly visualized. By multiplexing with a permeable rhodamine, we are able to differentiate extra- and intracellular SNAP- and Halo-tags, including those installed on the glucagon-like peptide-1 receptor, a prototypical class B G protein-coupled receptor. Sulfo549 and Sulfo646 also labelled transfected neurons derived from induced pluripotent stem cells (iPSCs), allowing STED nanoscopy of the axonal membrane. Together, this work provides a new avenue for rendering dyes impermeable for exclusive extracellular visualization via self-labelling protein tags. We anticipate that Sulfo549, Sulfo646 and their congeners will be useful for a number of cell biology applications where labelling of intracellular sites interferes with accurate surface protein analysis.
Date Issued
2022-08-14
Date Acceptance
2022-01-11
Citation
Organic and Biomolecular Chemistry, 2022, 20 (30), pp.5967-5980
ISSN
1477-0520
Publisher
Royal Society of Chemistry
Start Page
5967
End Page
5980
Journal / Book Title
Organic and Biomolecular Chemistry
Volume
20
Issue
30
Copyright Statement
This journal is © The Royal Society of Chemistry 2022 Open Access Article This article is licensed under a Creative Commons Attribution 3.0 Unported Licence.
License URL
Identifier
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000758357700001&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=a2bf6146997ec60c407a63945d4e92bb
Subjects
Chemistry
Chemistry, Organic
DYES
GENERAL-METHOD
LIVE-CELL
Physical Sciences
PROBES
Science & Technology
STED MICROSCOPY
SUPERRESOLUTION MICROSCOPY
Publication Status
Published
Coverage Spatial
England
Article Number
d1ob02216d
Date Publish Online
2022-02-21