Key Process and Factors Controlling the Direct Translocation of Cell-Penetrating Peptide through Bio-Membrane
Cell-penetrating peptide (CPP) can directly penetrate the cytosol (cytolysis) and is expected to be a potent vector for a drug delivery system (DDS). Although there is general agreement that CPP cytolysis is related to dynamic membrane deformation, a distinctive process has yet to be established. He...
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MDPI AG
2020-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/15/5466 |
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| author | Kazutami Sakamoto Taku Morishita Kenichi Aburai Kenichi Sakai Masahiko Abe Ikuhiko Nakase Shiroh Futaki Hideki Sakai |
| author_facet | Kazutami Sakamoto Taku Morishita Kenichi Aburai Kenichi Sakai Masahiko Abe Ikuhiko Nakase Shiroh Futaki Hideki Sakai |
| author_sort | Kazutami Sakamoto |
| collection | DOAJ |
| description | Cell-penetrating peptide (CPP) can directly penetrate the cytosol (cytolysis) and is expected to be a potent vector for a drug delivery system (DDS). Although there is general agreement that CPP cytolysis is related to dynamic membrane deformation, a distinctive process has yet to be established. Here, we report the key process and factors controlling CPP cytolysis. To elucidate the task, we have introduced trypsin digestion of adsorbed CPP onto giant unilamellar vesicle (GUV) to quantify the adsorption and internalization (cytolysis) separately. Also, the time-course analysis was introduced for the geometric calculation of adsorption and internalization amount per lipid molecule consisting of GUV. As a result, we found that adsorption and internalization assumed to occur successively by CPP molecule come into contact with membrane lipid. Adsorption is quick to saturate within 10 min, while cytolysis of each CPP on the membrane follows successively. After adsorption is saturated, cytolysis proceeds further linearly by time with a different rate constant that is dependent on the osmotic pressure. We also found that temperature and lipid composition influence cytolysis by modulating lipid mobility. The electrolyte in the outer media is also affected as a chemical mediator to control CPP cytolysis by following the Hoffmeister effect for membrane hydration. These results confirmed the mechanism of cytolysis as temporal and local phase transfer of membrane lipid from Lα to Mesh<sub>1</sub>, which has punctured bilayer morphologies. |
| first_indexed | 2024-03-10T18:05:02Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T18:05:02Z |
| publishDate | 2020-07-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-41879a4bd5dc48d4ace74e547b3087c32023-11-20T08:34:20ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-07-012115546610.3390/ijms21155466Key Process and Factors Controlling the Direct Translocation of Cell-Penetrating Peptide through Bio-MembraneKazutami Sakamoto0Taku Morishita1Kenichi Aburai2Kenichi Sakai3Masahiko Abe4Ikuhiko Nakase5Shiroh Futaki6Hideki Sakai7Department of Pure and Applied Chemistry, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, JapanDepartment of Pure and Applied Chemistry, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, JapanDepartment of Pure and Applied Chemistry, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, JapanDepartment of Pure and Applied Chemistry, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, JapanDepartment of Pure and Applied Chemistry, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, JapanNanoscience and Nanotechnology Research Center, Research Organization for the 21st Century, Osaka Prefecture University, Naka-ku, Sakai, Osaka 599-8570, JapanInstitute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, JapanDepartment of Pure and Applied Chemistry, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, JapanCell-penetrating peptide (CPP) can directly penetrate the cytosol (cytolysis) and is expected to be a potent vector for a drug delivery system (DDS). Although there is general agreement that CPP cytolysis is related to dynamic membrane deformation, a distinctive process has yet to be established. Here, we report the key process and factors controlling CPP cytolysis. To elucidate the task, we have introduced trypsin digestion of adsorbed CPP onto giant unilamellar vesicle (GUV) to quantify the adsorption and internalization (cytolysis) separately. Also, the time-course analysis was introduced for the geometric calculation of adsorption and internalization amount per lipid molecule consisting of GUV. As a result, we found that adsorption and internalization assumed to occur successively by CPP molecule come into contact with membrane lipid. Adsorption is quick to saturate within 10 min, while cytolysis of each CPP on the membrane follows successively. After adsorption is saturated, cytolysis proceeds further linearly by time with a different rate constant that is dependent on the osmotic pressure. We also found that temperature and lipid composition influence cytolysis by modulating lipid mobility. The electrolyte in the outer media is also affected as a chemical mediator to control CPP cytolysis by following the Hoffmeister effect for membrane hydration. These results confirmed the mechanism of cytolysis as temporal and local phase transfer of membrane lipid from Lα to Mesh<sub>1</sub>, which has punctured bilayer morphologies.https://www.mdpi.com/1422-0067/21/15/5466cell-penetrating peptide (CPP)direct permeation (cytolysis)giant unilamellar vesicle (GUV)CPP adsorptionFITC-octa arginine (FITC-R8) |
| spellingShingle | Kazutami Sakamoto Taku Morishita Kenichi Aburai Kenichi Sakai Masahiko Abe Ikuhiko Nakase Shiroh Futaki Hideki Sakai Key Process and Factors Controlling the Direct Translocation of Cell-Penetrating Peptide through Bio-Membrane International Journal of Molecular Sciences cell-penetrating peptide (CPP) direct permeation (cytolysis) giant unilamellar vesicle (GUV) CPP adsorption FITC-octa arginine (FITC-R8) |
| title | Key Process and Factors Controlling the Direct Translocation of Cell-Penetrating Peptide through Bio-Membrane |
| title_full | Key Process and Factors Controlling the Direct Translocation of Cell-Penetrating Peptide through Bio-Membrane |
| title_fullStr | Key Process and Factors Controlling the Direct Translocation of Cell-Penetrating Peptide through Bio-Membrane |
| title_full_unstemmed | Key Process and Factors Controlling the Direct Translocation of Cell-Penetrating Peptide through Bio-Membrane |
| title_short | Key Process and Factors Controlling the Direct Translocation of Cell-Penetrating Peptide through Bio-Membrane |
| title_sort | key process and factors controlling the direct translocation of cell penetrating peptide through bio membrane |
| topic | cell-penetrating peptide (CPP) direct permeation (cytolysis) giant unilamellar vesicle (GUV) CPP adsorption FITC-octa arginine (FITC-R8) |
| url | https://www.mdpi.com/1422-0067/21/15/5466 |
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