Molecular Mechanism of P53 Peptide Permeation through Lipid Membranes from Solid-State NMR Spectroscopy and Molecular Dynamics Simulations

Molecular Mechanism of P53 Peptide Permeation through Lipid Membranes from Solid-State NMR Spectroscopy and Molecular Dynamics Simulations

Journal of the American Chemical Society

10.1021/jacs.4c04230

https://doi.org/10.1021/jacs.4c04230

Mingyue Li, Jianguo Li, Xingyu Lu, Ryan Schroder, Arun Chandramohan, W. Peter Wuelfing, Allen C. Templeton, Wei Xu, Marian Gindy, Filippos Kesisoglou, Jing Ling, Tomi Sawyer, Chandra S. Verma, Anthony W. Partridge, Yongchao Su

Functional genomics

07 August 2024

Li, M., Li, J., Lu, X., Schroder, R., Chandramohan, A., Wuelfing, W. P., Templeton, A. C., Xu, W., Gindy, M., Kesisoglou, F., Ling, J., Sawyer, T., Verma, C. S., Partridge, A. W., & Su, Y. (2024). Molecular Mechanism of P53 Peptide Permeation through Lipid Membranes from Solid-State NMR Spectroscopy and Molecular Dynamics Simulations. Journal of the American Chemical Society, 146(33), 23075–23091. https://doi.org/10.1021/jacs.4c04230

Macrocyclic peptides show promise in targeting high-value therapeutically relevant binding sites due to their high affinity and specificity. However, their clinical application is often hindered by low membrane permeability, which limits their effectiveness against intracellular targets. Previous studies focused on peptide conformations in various solvents, leaving a gap in understanding their interactions with and translocation through lipid bilayers. Addressing this, our study explores the membrane interactions of stapled peptides, a subclass of macrocyclic peptides, using solid-state nuclear magnetic resonance (ssNMR) spectroscopy and molecular dynamics (MD) simulations. We conducted ssNMR measurements on ATSP-7041M, a prototypical stapled peptide, to understand its interaction with lipid membranes, leading to an MD-informed model for peptide membrane permeation. Our findings reveal that ATSP-7041M adopts a stable α-helical structure upon membrane binding, facilitated by a cation−π interaction between its phenylalanine side chain and the lipid headgroup. This interaction makes the membrane-bound state energetically favorable, facilitating membrane affinity and insertion. The bound peptide displayed asymmetric insertion depths, with the C-terminus penetrating deeper (approximately 9 Å) than the N-terminus (approximately 4.3 Å) relative to the lipid headgroups. Contrary to expectations, peptide dynamics was not hindered by membrane binding and exhibited rapid motions similar to cell-penetrating peptides. These dynamic interactions and peptide–lipid affinity appear to be crucial for membrane permeation. MD simulations indicated a thermodynamically stable transmembrane conformation of ATSP-7041M, reducing the energy barrier for translocation. Our study offers an in silico view of ATSP-7041M’s translocation from the extracellular to the intracellular region, highlighting the significance of peptide–lipid interactions and dynamics in enabling peptide transit through membranes.

Publisher Copyright

This research / project is supported by the Agency for Science, Technology and Research - Career Development Award
Grant Reference no. : C210112036

This document is the Accepted Manuscript version of a Published Work that appeared in final form in Journal of the American Chemical Society, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see 10.1021/jacs.4c04230.

https://oar.a-star.edu.sg/communities-collections/articles/22193

0002-7863
1520-5126

Bioinformatics Institute