Genetic, cellular, and structural characterization of the membrane potential-dependent cell-penetrating peptide translocation pore Article Swipe
Evgeniya Trofimenko
,
Gianvito Grasso
,
Mathieu Heulot
,
Nadja Chevalier
,
Marco A. Deriu
,
Gilles Dubuis
,
Yoan Arribat
,
Marc Serulla
,
Sébastien Michel
,
Gil Vantomme
,
Florine Ory
,
Linh Chi Dam
,
Julien Puyal
,
Francesca Amati
,
Anita Lüthi
,
Andrea Danani
,
Christian Widmann
·
YOU?
·
· 2021
· Open Access
·
· DOI: https://doi.org/10.7554/elife.69832
YOU?
·
· 2021
· Open Access
·
· DOI: https://doi.org/10.7554/elife.69832
Cell-penetrating peptides (CPPs) allow intracellular delivery of bioactive cargo molecules. The mechanisms allowing CPPs to enter cells are ill-defined. Using a CRISPR/Cas9-based screening, we discovered that KCNQ5, KCNN4, and KCNK5 potassium channels positively modulate cationic CPP direct translocation into cells by decreasing the transmembrane potential (V m ). These findings provide the first unbiased genetic validation of the role of V m in CPP translocation in cells. In silico modeling and live cell experiments indicate that CPPs, by bringing positive charges on the outer surface of the plasma membrane, decrease the V m to very low values (–150 mV or less), a situation we have coined megapolarization that then triggers formation of water pores used by CPPs to enter cells. Megapolarization lowers the free energy barrier associated with CPP membrane translocation. Using dyes of varying dimensions in CPP co-entry experiments, the diameter of the water pores in living cells was estimated to be 2 (–5) nm, in accordance with the structural characteristics of the pores predicted by in silico modeling. Pharmacological manipulation to lower transmembrane potential boosted CPP cellular internalization in zebrafish and mouse models. Besides identifying the first proteins that regulate CPP translocation, this work characterized key mechanistic steps used by CPPs to cross cellular membranes. This opens the ground for strategies aimed at improving the ability of cells to capture CPP-linked cargos in vitro and in vivo.
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Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.7554/elife.69832
- https://elifesciences.org/articles/69832.bib
- OA Status
- gold
- Cited By
- 52
- References
- 157
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W3208528933
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W3208528933Canonical identifier for this work in OpenAlex
- DOI
-
https://doi.org/10.7554/elife.69832Digital Object Identifier
- Title
-
Genetic, cellular, and structural characterization of the membrane potential-dependent cell-penetrating peptide translocation poreWork title
- Type
-
articleOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2021Year of publication
- Publication date
-
2021-10-29Full publication date if available
- Authors
-
Evgeniya Trofimenko, Gianvito Grasso, Mathieu Heulot, Nadja Chevalier, Marco A. Deriu, Gilles Dubuis, Yoan Arribat, Marc Serulla, Sébastien Michel, Gil Vantomme, Florine Ory, Linh Chi Dam, Julien Puyal, Francesca Amati, Anita Lüthi, Andrea Danani, Christian WidmannList of authors in order
- Landing page
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https://doi.org/10.7554/elife.69832Publisher landing page
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https://elifesciences.org/articles/69832.bibDirect link to full text PDF
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YesWhether a free full text is available
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goldOpen access status per OpenAlex
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https://elifesciences.org/articles/69832.bibDirect OA link when available
- Concepts
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Chromosomal translocation, Characterization (materials science), Cell biology, Peptide, Cell-penetrating peptide, Membrane, Biophysics, Cell, Chemistry, Biology, Structural biology, Computational biology, Biochemistry, Nanotechnology, Materials science, GeneTop concepts (fields/topics) attached by OpenAlex
- Cited by
-
52Total citation count in OpenAlex
- Citations by year (recent)
-
2025: 12, 2024: 12, 2023: 12, 2022: 14, 2021: 2Per-year citation counts (last 5 years)
- References (count)
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157Number of works referenced by this work
- Related works (count)
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10Other works algorithmically related by OpenAlex
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