Puromycin Analogues Capable of Multiplexed Imaging and Profiling of Protein Synthesis and Dynamics in Live Cells and Neurons |
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| Authors: | Dr. Jingyan Ge Dr. Cheng‐Wu Zhang Xue Wen Ng Bo Peng Sijun Pan Shubo Du Danyang Wang Prof. Dr. Lin Li Dr. Kah‐Leong Lim Prof. Dr. Thorsten Wohland Prof. Dr. Shao Q. Yao |
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| Affiliation: | 1. Department of Chemistry, National University of Singapore, Singapore, Singapore;2. Institute of Bioengineering, Zhejiang University of Technology, Hangzhou, China;3. Key Laboratory of Flexible Electronics & Institute of Advanced Materials, Nanjing Tech University, Nanjing, China;4. National Neuroscience Institute, Singapore, Singapore;5. Department of Biological Sciences, National University of Singapore, Singapore |
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| Abstract: | Newly synthesized proteins constitute an important subset of the proteome involved in every cellular process, yet existing chemical tools used to study them have major shortcomings. Herein we report a suite of cell‐permeable puromycin analogues capable of being metabolically incorporated into newly synthesized proteins in different mammalian cells, including neuronal cells. Subsequent labeling with suitable bioorthogonal reporters, in both fixed and live cells, enabled direct imaging and enrichment of these proteins. By taking advantage of the mutually orthogonal reactivity of these analogues, we showed multiplexed labeling of different protein populations, as well as quantitative measurements of protein dynamics by fluorescence correlation spectroscopy, could be achieved in live‐cell environments. |
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| Keywords: | bioorthogonality metabolic incorporation multiplexed imaging newly synthesized proteins puromycin |
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