Structural flexibility of apolipoprotein E-derived arginine-rich peptides improves their cell penetration capability
© 2023. The Author(s)..
Amphipathic arginine-rich peptide, A2-17, exhibits moderate perturbation of lipid membranes and the highest cell penetration among its structural isomers. We investigated the direct cell-membrane penetration mechanism of the A2-17 peptide while focusing on structural flexibility. We designed conformationally constrained versions of A2-17, stapled (StpA2-17) and stitched (StchA2-17), whose α-helical conformations were stabilized by chemical crosslinking. Circular dichroism confirmed that StpA2-17 and StchA2-17 had higher α-helix content than A2-17 in aqueous solution. Upon liposome binding, only A2-17 exhibited a coil-to-helix transition. Confocal microscopy revealed that A2-17 had higher cell penetration efficiency than StpA2-17, whereas StchA2-17 remained on the cell membrane without cell penetration. Although the tryptophan fluorescence analysis suggested that A2-17 and its analogs had similar membrane-insertion positions between the interface and hydrophobic core, StchA2-17 exhibited a higher membrane affinity than A2-17 or StpA2-17. Atomic force microscopy demonstrated that A2-17 reduced the mechanical rigidity of liposomes to a greater extent than StpA2-17 and StchA2-17. Finally, electrophysiological analysis showed that A2-17 induced a higher charge influx through transient pores in a planer lipid bilayer than StpA2-17 and StchA2-17. These findings indicate that structural flexibility, which enables diverse conformations of A2-17, leads to a membrane perturbation mode that contributes to cell membrane penetration.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:13 |
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| Enthalten in: |
Scientific reports - 13(2023), 1 vom: 08. Nov., Seite 19396 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Takechi-Haraya, Yuki [VerfasserIn] |
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| Links: |
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| Themen: |
94ZLA3W45F |
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| Anmerkungen: |
Date Completed 09.11.2023 Date Revised 11.11.2023 published: Electronic Citation Status MEDLINE |
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| doi: |
10.1038/s41598-023-46754-0 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM364303719 |
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| 100 | 1 | |a Takechi-Haraya, Yuki |e verfasserin |4 aut | |
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| 520 | |a Amphipathic arginine-rich peptide, A2-17, exhibits moderate perturbation of lipid membranes and the highest cell penetration among its structural isomers. We investigated the direct cell-membrane penetration mechanism of the A2-17 peptide while focusing on structural flexibility. We designed conformationally constrained versions of A2-17, stapled (StpA2-17) and stitched (StchA2-17), whose α-helical conformations were stabilized by chemical crosslinking. Circular dichroism confirmed that StpA2-17 and StchA2-17 had higher α-helix content than A2-17 in aqueous solution. Upon liposome binding, only A2-17 exhibited a coil-to-helix transition. Confocal microscopy revealed that A2-17 had higher cell penetration efficiency than StpA2-17, whereas StchA2-17 remained on the cell membrane without cell penetration. Although the tryptophan fluorescence analysis suggested that A2-17 and its analogs had similar membrane-insertion positions between the interface and hydrophobic core, StchA2-17 exhibited a higher membrane affinity than A2-17 or StpA2-17. Atomic force microscopy demonstrated that A2-17 reduced the mechanical rigidity of liposomes to a greater extent than StpA2-17 and StchA2-17. Finally, electrophysiological analysis showed that A2-17 induced a higher charge influx through transient pores in a planer lipid bilayer than StpA2-17 and StchA2-17. These findings indicate that structural flexibility, which enables diverse conformations of A2-17, leads to a membrane perturbation mode that contributes to cell membrane penetration | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Arginine |2 NLM | |
| 650 | 7 | |a 94ZLA3W45F |2 NLM | |
| 650 | 7 | |a Liposomes |2 NLM | |
| 650 | 7 | |a Peptides |2 NLM | |
| 650 | 7 | |a Apolipoproteins E |2 NLM | |
| 700 | 1 | |a Ohgita, Takashi |e verfasserin |4 aut | |
| 700 | 1 | |a Usui, Akiko |e verfasserin |4 aut | |
| 700 | 1 | |a Nishitsuji, Kazuchika |e verfasserin |4 aut | |
| 700 | 1 | |a Uchimura, Kenji |e verfasserin |4 aut | |
| 700 | 1 | |a Abe, Yasuhiro |e verfasserin |4 aut | |
| 700 | 1 | |a Kawano, Ryuji |e verfasserin |4 aut | |
| 700 | 1 | |a Konaklieva, Monika I |e verfasserin |4 aut | |
| 700 | 1 | |a Reimund, Mart |e verfasserin |4 aut | |
| 700 | 1 | |a Remaley, Alan T |e verfasserin |4 aut | |
| 700 | 1 | |a Sato, Yoji |e verfasserin |4 aut | |
| 700 | 1 | |a Izutsu, Ken-Ichi |e verfasserin |4 aut | |
| 700 | 1 | |a Saito, Hiroyuki |e verfasserin |4 aut | |
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