Covalently Linked Peptides and Membrane Potential Enable CyaA Segment Translocation
Abstract
The adenylate cyclase toxin (CyaA) from Bordetella pertussis intoxicates host cells by directly translocating its N‐terminal catalytic domain across the plasma membrane; however, the forces driving this unique process remain poorly defined. Here, we dissect the membrane translocation mechanisms of two peptide segments derived from CyaA: P233 and P454 from the catalytic domain and the translocation region, respectively. Both P454 and P233 are calmodulin‐binding segments that are sequentially involved in the translocation and activation of the catalytic domain. Using a newly developed Droplet Interface Bilayer (DIB) approach, called DIB‐Pipette, which enables direct visualization of peptide transport under controlled membrane potentials, we show that P454 translocates across membranes independently of membrane potential, whereas P233 translocation requires a negative electric membrane potential. Strikingly, covalent coupling of P233 and P454 enables efficient translocation of the resulting peptide even in the absence of a membrane potential. Together, these results suggest that two distinct membrane‐active segments within CyaA act cooperatively to promote translocation at the peptide level, revealing an intrinsic mechanism that may contribute to membrane potential‐dependent translocation. These findings provide new mechanistic insights into the CyaA cell intoxication process and reveal a multifunctional strategy for protein delivery across membranes.
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BibTeX
@article{scilironi2026covalently,
title = {{Covalently Linked Peptides and Membrane Potential Enable CyaA Segment Translocation}},
author = {Gaia Scilironi and Nicolas Carvalho and Jacinthe Frangieh and Corentin Léger and Dorothée Raoux-Barbot and J. Iñaki Guijarro and Daniel Ladant and Sophie Cribier and Nicolas Rodriguez and Alexandre Chenal},
journal = {Advanced Science},
year = {2026},
month = aug,
doi = {10.1002/advs.77133},
url = {https://hal.science/hal-05783983v2},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Covalently Linked Peptides and Membrane Potential Enable CyaA Segment Translocation
AU - Gaia Scilironi
AU - Nicolas Carvalho
AU - Jacinthe Frangieh
AU - Corentin Léger
AU - Dorothée Raoux-Barbot
AU - J. Iñaki Guijarro
AU - Daniel Ladant
AU - Sophie Cribier
AU - Nicolas Rodriguez
AU - Alexandre Chenal
PY - 2026
DA - 2026/08/21
JO - Advanced Science
DO - 10.1002/advs.77133
UR - https://hal.science/hal-05783983v2
AB - The adenylate cyclase toxin (CyaA) from Bordetella pertussis intoxicates host cells by directly translocating its N‐terminal catalytic domain across the plasma membrane; however, the forces driving this unique process remain poorly defined. Here, we dissect the membrane translocation mechanisms of two peptide segments derived from CyaA: P233 and P454 from the catalytic domain and the translocation region, respectively. Both P454 and P233 are calmodulin‐binding segments that are sequentially involved in the translocation and activation of the catalytic domain. Using a newly developed Droplet Interface Bilayer (DIB) approach, called DIB‐Pipette, which enables direct visualization of peptide transport under controlled membrane potentials, we show that P454 translocates across membranes independently of membrane potential, whereas P233 translocation requires a negative electric membrane potential. Strikingly, covalent coupling of P233 and P454 enables efficient translocation of the resulting peptide even in the absence of a membrane potential. Together, these results suggest that two distinct membrane‐active segments within CyaA act cooperatively to promote translocation at the peptide level, revealing an intrinsic mechanism that may contribute to membrane potential‐dependent translocation. These findings provide new mechanistic insights into the CyaA cell intoxication process and reveal a multifunctional strategy for protein delivery across membranes.
SN - 2198-3844
ER -