دورية أكاديمية
Spatiotemporally Controlled Photolabeling of Genetically Unmodified Proteins in Live Cells
| العنوان: | Spatiotemporally Controlled Photolabeling of Genetically Unmodified Proteins in Live Cells |
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| المؤلفون: | Huaibin Yu, Shuangshuang Wang, Yu Fu, Manfred Wagner, Tanja Weil, Shaoqin Liu, Weining Zhao, Fangrui Zhong, Yuzhou Wu |
| سنة النشر: | 2024 |
| مصطلحات موضوعية: | Biophysics, Biochemistry, Cell Biology, Genetics, Molecular Biology, Physiology, Pharmacology, Biotechnology, Immunology, Cancer, Inorganic Chemistry, Virology, Computational Biology, Chemical Sciences not elsewhere classified, Physical Sciences not elsewhere classified, spatiotemporally controlled photolabeling, spatiotemporally controlled installation, proximity coupling reaction, peptide short chain, g ., biotin, discovering dynamic functions, directed photoclick strategy, >- naphthoquinone methide, spatiotemporally controlled labeling, genetically unmodified proteins, understanding protein functions, resolved labeling, endogenous proteins, via acylation |
| الوصف: | Selective labeling of the protein of interest (POI) in genetically unmodified live cells is crucial for understanding protein functions and kinetics in their natural habitat. In particular, spatiotemporally controlled installation of the labels on a POI under light control without affecting their original activity is in high demand but is a tremendous challenge. Here, we describe a novel ligand-directed photoclick strategy for spatiotemporally controlled labeling of endogenous proteins in live cells. It was realized with a designer labeling reagent skillfully integrating the photochemistries of 2-nitrophenylpropyloxycarbonyl and 3-hydroxymethyl-2-naphthol with an affinity ligand. Highly electrophilic ortho -naphthoquinone methide was photochemically released and underwent a proximity coupling reaction with nucleophilic amino acid residues on the POI in live cells. With fluorescein as a marker, this photoclick strategy enables time-resolved labeling of carbonic anhydrase subtypes localized either on the cell membrane or in the cytoplasm and a discriminable visualization of their metabolic kinetics. Given the versatility underlined by facilely tethering other functional entities (e.g., biotin, a peptide short chain) via acylation or (in cell) Huisgen cycloaddition, this affinity-driven photoclick chemistry opens up enormous opportunities for discovering dynamic functions and mechanistic interrogation of endogenous proteins in live cells. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | unknown |
| العلاقة: | https://figshare.com/articles/journal_contribution/Spatiotemporally_Controlled_Photolabeling_of_Genetically_Unmodified_Proteins_in_Live_Cells/25033815Test |
| DOI: | 10.1021/acs.analchem.3c04099.s001 |
| الإتاحة: | https://doi.org/10.1021/acs.analchem.3c04099.s001Test https://figshare.com/articles/journal_contribution/Spatiotemporally_Controlled_Photolabeling_of_Genetically_Unmodified_Proteins_in_Live_Cells/25033815Test |
| حقوق: | CC BY-NC 4.0 |
| رقم الانضمام: | edsbas.E88ACF08 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1021/acs.analchem.3c04099.s001 |
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