Reference - Detail
| RRC ID | 63167 |
|---|---|
| Author | Kim H, Watanabe S, Kitamatsu M, Watanabe K, Ohtsuki T. |
| Title | Cell cycle dependence of apoptosis photo-triggered using peptide-photosensitizer conjugate. |
| Journal | Sci Rep |
| Abstract |
Investigation of the relevance between cell cycle status and the bioactivity of exogenously delivered biomacromolecules is hindered by their time-consuming cell internalization and the cytotoxicity of transfection methods. In this study, we addressed these problems by utilizing the photochemical internalization (PCI) method using a peptide/protein-photosensitizer conjugate, which enables immediate cytoplasmic internalization of the bioactive peptides/proteins in a light-dependent manner with low cytotoxicity. To identify the cell-cycle dependent apoptosis, a TatBim peptide-photosensitizer conjugate (TatBim-PS) with apoptotic activity was photo-dependently internalized into HeLa cells expressing a fluorescent ubiquitination-based cell cycle indicator (Fucci2). Upon irradiation, cytoplasmic TatBim-PS internalization exceeded 95% for all cells classified in the G1, S, and G2/M cell cycle phases with no significant differences between groups. TatBim-PS-mediated apoptosis was more efficiently triggered by photoirradiation in the G1/S transition than in the G1 and S/G2/M phases, suggesting high sensitivity of the former phase to Bim-induced apoptosis. Thus, the cell cycle dependence of Bim peptide-induced apoptosis was successfully investigated using Fucci2 indicator and the PCI method. Since PCI-mediated cytoplasmic internalization of peptides is rapid and does not span multiple cell cycle phases, the Fucci-PCI method constitutes a promising tool for analyzing the cell cycle dependence of peptides/protein functions. |
| Volume | 10(1) |
| Pages | 19087 |
| Published | 2020-11-5 |
| DOI | 10.1038/s41598-020-76100-7 |
| PII | 10.1038/s41598-020-76100-7 |
| PMID | 33154435 |
| PMC | PMC7644668 |
| MeSH | Amino Acid Sequence Apoptosis / drug effects* Bcl-2-Like Protein 11 / chemistry Bcl-2-Like Protein 11 / pharmacokinetics Bcl-2-Like Protein 11 / pharmacology Cell Cycle / drug effects* Cytoplasm / metabolism Fluorescent Dyes HeLa Cells Humans Microscopy, Fluorescence Peptides / chemistry Peptides / pharmacokinetics Peptides / pharmacology* Photosensitizing Agents / chemistry Photosensitizing Agents / pharmacokinetics Photosensitizing Agents / pharmacology* Thymidine / pharmacology |
| IF | 3.998 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 1 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | HeLa/Fucci2(RCB2867) |