Cell-penetrating Peptides

By Jean-Philippe Richard, K. Melikov, E. Vivés, Corinne Ramos, B. Verbeure, M. Gait, L. Chernomordik, B. Lebleu. Journal of Biological Chemistry, 2003

1,123 citations recorded by the source corpus.

Cellular uptake of a family of cationic cell-penetrating peptides (examples include Tat peptides and penetratin) have been ascribed in the literature to a mechanism that does not involve endocytosis. In this work we reevaluate the mechanisms of cellular uptake of Tat 48–60 and (Arg)9. We demonstrate here that cell fixation, even in mild conditions, leads to the artifactual uptake of these peptides. Moreover, we show that flow cytometry analysis cannot be used validly to evaluate cellular uptake unless a step of trypsin digestion of the cell membrane-adsorbed peptide is included in the protocol. Fluorescence microscopy on live unfixed cells shows characteristic endosomal distribution of peptides. Flow cytometry analysis indicates that the kinetics of uptake are similar to the kinetics of endocytosis. Peptide uptake is inhibited by incubation at low temperature and cellular ATP pool depletion. Similar data were obtained for Tat-conjugated peptide nucleic acids. These data are consistent with the involvement of endocytosis in the cellular internalization of cell-penetrating peptides and their conjugates to peptide nucleic acids.

Subfields: Immunology, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/d5aaf0933d18e1f3c269eda4afd69e5ea8ed1d20

How researchers see this paper

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In Immunology

1. D. Begley

In Molecular Biology

1. I. Kaplan
2. J. Wadia
3. S. Dowdy
4. D. Begley
5. P. Gleeson
6. J. P. Lim

Citations in the corpus

• Cellular uptake of arginine-rich peptides: roles for macropinocytosis and actin rearrangement. (2004, background): “Several endocytic routes have been suggested for the cellular uptake of Tat and other arginine peptides [7,19– 21,32].”
• Delivery of therapeutic agents to the central nervous system: the problems and the possibilities. (2004, result): “Other studies suggest that cell-penetrating peptide may induce an endocytic event at the plasma membrane perhaps with a similar mechanism to that of AMT (Green et al., 2003; Richard et al., 2003).”
• Cationic TAT peptide transduction domain enters cells by macropinocytosis. (2005, result): “However, recent studies have questioned these early observations and demonstrated that PTD peptides bound to cell surface become redistributed during cell fixation resulting in inaccurate measurements [15].”