Using Fluorescence On/Off to Trace Tandem Nanofiber Assembly/Disassembly in Living Cells
| Autor: | Miaomiao Zhang, Huae Xu, Chenchen Wang, Gaolin Liang, Chen Yang, Haisi Wu |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
biology
Tandem Chemistry 010401 analytical chemistry Nanofibers Living cell 010402 general chemistry biology.organism_classification 01 natural sciences Fluorescence In vitro 0104 chemical sciences Analytical Chemistry HeLa Cell outer membrane Assembly disassembly Nanofiber Biophysics Humans Fluorescent Dyes HeLa Cells |
| Zdroj: | Analytical Chemistry. 93:5665-5669 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.1c00220 |
| Popis: | To track an intact biological process inside cells, continuous showing of the assembly/disassembly process is needed and fluorescence is advantageous in characterizing these processes. However, using fluorescence "on/off" to observe a sequential assembly/disassembly process in living cells has not been reported. Herein, we rationally designed a probe PEA-NBD-Yp and employed its fluorescence "on/off" to trace tandem assembly/disassembly of nanofibers in living HeLa cells. In vitro experiments validated that PEA-NBD-Yp could be efficiently dephosphorylated by ALP to yield PEA-NBD-Y, which self-assembled into nanofibers with the NBD fluorescence "on". Also, the PEA-NBD-Y nanofiber was disassembled by GSH, accompanied by fluorescence "off". Living cell imaging (together with ALP-inhibition or GSH-blocking) experiments sequentially showed the self-assembling nanofibers on the cell outer membrane with fluorescence "on" (On1), translocated inside cells (On2), and disassembled by GSH with fluorescence "off" (Off2). We anticipate that our strategy of one probe conferring temporal "on/off" fluorescence signals might provide people with a new tool to deeply understand a biological event in living cells in the near future. |
| Databáze: | OpenAIRE |
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