期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 61, 期 47, 页码 -出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202207551
关键词
Cell-Penetrating Peptides; Chemical Protein Synthesis; Fluorescence; Intracellular Protein Delivery; Nucleation Zones
资金
- German-Israeli Foundation for Scientific Research and Development [I-1488-302.5/2019]
- DFG [RTG2473, 392923329]
- Alexander von Humboldt Fellowship
- Margarita Salas Fellowship from Spanish Ministry of Universities
By studying DABCYL-modified cR10 peptides and their derivatives, it was found that the efficiency of Ub cargo entering living cells can be significantly improved, which is of great research value.
Modifying cyclic cell-penetrating deca-arginine (cR10) peptides with 4-(4-dimethylaminophenylazo)benzoic acid (DABCYL) improves the uptake efficiency of synthetic ubiquitin (Ub) cargoes into living cells. To probe the role of the DABCYL moiety, we performed time-lapse microscopy and fluorescence lifetime imaging microscopy (FLIM) of fluorescent DABCYL-R10 to evaluate the impact on cell entry by the formation of nucleation zones. Furthermore, we performed a structure-uptake relationship study with 13 DABCYL derivatives coupled to CPP to examine their effect on the cell-uptake efficiency when conjugated to mono-Ub through disulfide linkages. Our results show that through structure variations of the DABCYL moiety alone we could reach, at nanomolar concentration, an additional threefold increase in the cytosolic delivery of Ub, which will enable studies on various intracellular processes related to Ub signaling.
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