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- W3131286983 abstract "Protein delivery presents an important topic of interest due to its therapeutic and scientific applications. Majority of the delivery methods employ either protein modification to ensure protein penetration inside the cell or nanocarrier delivery with following release in the cytoplasm. Functional protein uptake by the adherent cells from the medium without additional modifications may prove to be a valuable scientific tool.Here we report unexpected effect of the puromycin treatment on the HEK293 cells. Puromycin is typically used to abruptly finish protein translation on ribosomes by releasing the polypeptide chain. After the prolonged (3 h+) puromycin exposure cell import protein from the medium. We have used the mitochondrially targeted MTS-SNAP-tag protein and the protein was later found in the mitochondria of the cell by fluorescence microscopy. At the same time, no endosomal structures were observed, suggesting that this import mechanism either bypasses endocytosis or is caused by membrane becoming porous. Such an effect was reported previously for unusually high puromycin concentrations, but not for a prolonged exposure in the typically used 20 µg/ml concentration. Similar effect was not seen for our fluorescent protein in the untreated or cycloheximide-treated cells. This suggests that the puromycin influence is not directly connected to either the nature of the protein used or the translation stoppage as is the case with cycloheximide. Overall, we believe that this effect on the protein uptake warrants further investigations and, if it works with other proteins, may provide a useful tool to deliver protein into the cells.This work was supported by RFBR (No. 20-54-12010) and DFG (No. HA2868/14-1). VB, AM, IO are thankful to the Ministry of Science and Higher Education of the Russian Federation (agreement No. 075-00337-20-03, project FSMG −2020-0003)." @default.
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- W3131286983 date "2021-02-01" @default.
- W3131286983 modified "2023-10-15" @default.
- W3131286983 title "Puromycin Causes Protein Uptake by Cells from the Medium" @default.
- W3131286983 doi "https://doi.org/10.1016/j.bpj.2020.11.1102" @default.
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