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  Efflux pump insensitive rhodamine–jasplakinolide conjugates for G- and F-actin imaging in living cells

Gerasimaite, R., Seikowski, J., Schimpfhauser, J., Kostiuk, G., Gilat, T., D'Este, E., et al. (2020). Efflux pump insensitive rhodamine–jasplakinolide conjugates for G- and F-actin imaging in living cells. Organic Biomolecular Chemistry, 18(15), 2929-2937. doi:10.1039/D0OB00369G.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0006-9F1F-6 Version Permalink: http://hdl.handle.net/21.11116/0000-0006-9F21-2
Genre: Journal Article

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 Creators:
Gerasimaite, R.1, Author              
Seikowski, J., Author
Schimpfhauser, J., Author
Kostiuk, G., Author
Gilat, T.2, Author              
D'Este, E., Author
Schnorrenberg, S.2, Author              
Lukinavicius, G.1, Author              
Affiliations:
1Laboratory of Chromatin Labeling and Imaging, Max Planck Institute for Biophysical Chemistry, Max Planck Society, ou_2616691              
2Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society, ou_578627              

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 Abstract: The actin cytoskeleton is crucial for endocytosis, intracellular trafficking, cell shape maintenance and a wide range of other cellular functions. Recently introduced cell-permeable fluorescent actin probes, such as SiR-actin, suffer from poor membrane permeability and stain some cell populations inhomogeneously due to the active efflux by the plasma membrane pumps. We analyzed a series of new probes composed of jasplakinolide and modified rhodamine fluorophores and found that rhodamine positional isomerism has a profound effect on probe performance. The probes based on the 6'-carboxy-carbopyronine scaffold are considerably less susceptible to efflux and allow efficient staining without efflux pump inhibitors. They can be used for 2D and 3D fluorescence nanoscopy at high nanomolar concentrations without significant cytotoxicity. We show that jasplakinolide-based fluorescent probes bind not only to actin filaments, but also to G-actin, which enables imaging highly dynamic actin structures. We demonstrate an excellent performance of the new probes in multiple organisms and cell types: human cell lines, frog erythrocytes, fruit fly tissues and primary neurons.

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Language(s): eng - English
 Dates: 2020-03-262020-04-21
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1039/D0OB00369G
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Title: Organic Biomolecular Chemistry
Source Genre: Journal
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Pages: - Volume / Issue: 18 (15) Sequence Number: - Start / End Page: 2929 - 2937 Identifier: -