Ter-Avetisyan, Gohar ; Tünnemann, Gisela ; Nowak, Danny ; Nitschke, Matthias ; Herrmann, Andreas ; Drab, Marek ; Cardoso, M. Cristina (2009)
Cell entry of arginine-rich peptides is independent of endocytosis.
In: The Journal of biological chemistry, 284 (6)
Artikel, Bibliographie
Kurzbeschreibung (Abstract)
Arginine-rich peptides are a subclass of cell-penetrating peptides that are taken up by living cells and can be detected freely diffusing inside the cytoplasm and nucleoplasm. This phenomenon has been attributed to either an endocytic mode of uptake and a subsequent release from vesicles or to direct membrane penetration (transduction). To distinguish between both possibilities, we have blocked endocytic pathways suggested to be involved in uptake of cell-penetrating peptides. We have then monitored by confocal microscopy the uptake and distribution of the cell-penetrating transactivator of transcription (TAT) peptide into living mammalian cells over time. To prevent side effects of chemical inhibitors, we used genetically engineered cells as well as different temperature. We found that a knockdown of clathrin-mediated endocytosis and a knock-out of caveolin-mediated endocytosis did not affect the ability of TAT to enter cells. In addition, the TAT peptide showed the same intracellular distribution throughout the cytoplasm and nucleus as in control cells. Even incubation of cells at 4 degrees C did not abrogate TAT uptake nor change its intracellular distribution. We therefore conclude that this distribution results from TAT peptide that directly penetrated (transduced) the plasma membrane. The formation of nonselective pores is unlikely, because simultaneously added fluorophores were not taken up together with the TAT peptide. In summary, although the frequency and kinetics of TAT transduction varied between cell types, it was independent of endocytosis.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2009 |
| Autor(en): | Ter-Avetisyan, Gohar ; Tünnemann, Gisela ; Nowak, Danny ; Nitschke, Matthias ; Herrmann, Andreas ; Drab, Marek ; Cardoso, M. Cristina |
| Art des Eintrags: | Bibliographie |
| Titel: | Cell entry of arginine-rich peptides is independent of endocytosis. |
| Sprache: | Englisch |
| Publikationsjahr: | 2009 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | The Journal of biological chemistry |
| Jahrgang/Volume einer Zeitschrift: | 284 |
| (Heft-)Nummer: | 6 |
| URL / URN: | http://www.cardoso-lab.org/publications/Ter-Avetisyan_2009.p... |
| Zugehörige Links: | |
| Kurzbeschreibung (Abstract): | Arginine-rich peptides are a subclass of cell-penetrating peptides that are taken up by living cells and can be detected freely diffusing inside the cytoplasm and nucleoplasm. This phenomenon has been attributed to either an endocytic mode of uptake and a subsequent release from vesicles or to direct membrane penetration (transduction). To distinguish between both possibilities, we have blocked endocytic pathways suggested to be involved in uptake of cell-penetrating peptides. We have then monitored by confocal microscopy the uptake and distribution of the cell-penetrating transactivator of transcription (TAT) peptide into living mammalian cells over time. To prevent side effects of chemical inhibitors, we used genetically engineered cells as well as different temperature. We found that a knockdown of clathrin-mediated endocytosis and a knock-out of caveolin-mediated endocytosis did not affect the ability of TAT to enter cells. In addition, the TAT peptide showed the same intracellular distribution throughout the cytoplasm and nucleus as in control cells. Even incubation of cells at 4 degrees C did not abrogate TAT uptake nor change its intracellular distribution. We therefore conclude that this distribution results from TAT peptide that directly penetrated (transduced) the plasma membrane. The formation of nonselective pores is unlikely, because simultaneously added fluorophores were not taken up together with the TAT peptide. In summary, although the frequency and kinetics of TAT transduction varied between cell types, it was independent of endocytosis. |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie > Cell Biology and Epigenetics ?? fb10_zoologie ?? 10 Fachbereich Biologie |
| Hinterlegungsdatum: | 06 Mär 2010 16:20 |
| Letzte Änderung: | 05 Mär 2013 09:32 |
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