Transient pores account for cell-penetrating peptide and homeoprotein translocation
Abstract
Homeoproteins (HPs) and cell-penetrating peptides (CPPs) enter cells by endocytosis and direct membrane crossing (translocation). However, unlike endocytosis, translocation remains globally unknown. Here, we developed an electrophysiological approach to assess the internalization of CPPs (Tat, R 9 , penetratin and R 6 W 3 ) and the HPs Otx2 and En2 though single-cell unitary transient currents in mammalian cells. At resting membrane potential, CPPs or HPs lead to submillisecond transient pores, faster than any endocytosis event, which reveal the rapid passage of the peptide across the membrane i.e., by translocation. We evidenced that expression of specific membrane glycosaminoglycans is mandatory for translocation-induced transient pores. Associated transient currents are supralinearly enhanced by hyperpolarization and poorly affected by depolarization. Moreover, a CPP-conjugated bioactive cargo similarly translocates into cytosol. Finally, we show similar HPs-evoked transient pores in brain cortical pyramidal cells, showing the physiological relevance of translocation, with crucial biotechnological and therapeutical consequences for cell delivery purposes.
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BibTeX
@article{trofimenko2026transient,
title = {{Transient pores account for cell-penetrating peptide and homeoprotein translocation}},
author = {Evgeniya Trofimenko and Nicolas Gervasi and Sylvie Perez and Nicolas Rodriguez and Delphine Ravault and Sophie Cribier and Hugues Berry and Laurent Venance and Sandrine Sagan},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
year = {2026},
month = jun,
doi = {10.1073/pnas.2602649123},
url = {https://hal.science/hal-05746396v1},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Transient pores account for cell-penetrating peptide and homeoprotein translocation
AU - Evgeniya Trofimenko
AU - Nicolas Gervasi
AU - Sylvie Perez
AU - Nicolas Rodriguez
AU - Delphine Ravault
AU - Sophie Cribier
AU - Hugues Berry
AU - Laurent Venance
AU - Sandrine Sagan
PY - 2026
DA - 2026/06/22
JO - Proceedings of the National Academy of Sciences of the United States of America
DO - 10.1073/pnas.2602649123
UR - https://hal.science/hal-05746396v1
AB - Homeoproteins (HPs) and cell-penetrating peptides (CPPs) enter cells by endocytosis and direct membrane crossing (translocation). However, unlike endocytosis, translocation remains globally unknown. Here, we developed an electrophysiological approach to assess the internalization of CPPs (Tat, R 9 , penetratin and R 6 W 3 ) and the HPs Otx2 and En2 though single-cell unitary transient currents in mammalian cells. At resting membrane potential, CPPs or HPs lead to submillisecond transient pores, faster than any endocytosis event, which reveal the rapid passage of the peptide across the membrane i.e., by translocation. We evidenced that expression of specific membrane glycosaminoglycans is mandatory for translocation-induced transient pores. Associated transient currents are supralinearly enhanced by hyperpolarization and poorly affected by depolarization. Moreover, a CPP-conjugated bioactive cargo similarly translocates into cytosol. Finally, we show similar HPs-evoked transient pores in brain cortical pyramidal cells, showing the physiological relevance of translocation, with crucial biotechnological and therapeutical consequences for cell delivery purposes.
SN - 0027-8424
SN - 1091-6490
ER -