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- W2013928131 abstract "We present a combined experimental and theoretical study of the interactions between cell-penetrating peptides (CPPs) and lipid bilayers using dynamic AFM measurements. Understanding how CPPs can pass through cell membranes is critical for designing drug delivery agents. While CPPs like HIV-TAT have been widely studied, their ability to penetrate membranes directly, without active transport, is still a matter of considerable debate. Here, we directly measure TAT-lipid mechanics during the actual membrane translocation event using TAT-functionalized AFM probes to penetrate through a stack of lipid bilayers. Dynamic force spectroscopy revealed that both the bilayer breakthrough force and bilayer thickness depended on the TAT-bilayer contact time. The results provide a detailed view of how TAT interacts with the bilayer. Upon contact, TAT inserts into the bilayer headgroup to a depth of ∼1nm in ∼0.5ms. During this same timescale the bilayer thins from 4.2 to 3.2 nm, likely reflecting TAT-driven reorganization. Unexpectedly, the energy barrier for penetration actually increases 52 kT after the bilayer thins, indicating the new conformation has not weakened the bilayer and suggesting TAT cannot penetrate unassisted. Controls with polylysine or mercaptoundecanoic acid do not show this thinning behavior, implying the unique TAT-lipid interactions play a significant role. This approach can be further generalized to the interaction of a variety of peptides or membrane-active molecules with lipid bilayers. Theoretical modeling of dynamic force spectroscopy measurements shows distinct features related to the timescale and disruption of lipid membranes upon contact with a molecular species. By adapting existing AFM protocols to look for these features it will be possible to directly characterize, with microsecond resolution, the dynamics and energetics of the interactions between lipid bilayers and a wide range of peptides and molecular species." @default.
- W2013928131 created "2016-06-24" @default.
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- W2013928131 date "2010-01-01" @default.
- W2013928131 modified "2023-09-26" @default.
- W2013928131 title "AFM Force Spectroscopy on TAT Membrane Penetration" @default.
- W2013928131 doi "https://doi.org/10.1016/j.bpj.2009.12.1173" @default.
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