Improved Stabilities of Labeling Probes for the Selective Modification of Endogenous Proteins in Living Cells and In Vivo
| Autor: | Xin-Hui Lin, Yu-Chun Lin, Syuan-Yun Fan, Jui-Ping Li, Kuan-Yu Lin, Boon Khoon Tee, Jen-Kun Chen, Jung-I Hsu, Nitesh K. Gupta, Chak Hin Lam, Kui-Thong Tan |
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| Rok vydání: | 2021 |
| Předmět: |
Injections
Subcutaneous Cell Mice Nude Endogeny 010402 general chemistry Ligands 01 natural sciences Biochemistry Carbonic Anhydrase II Mice In vivo Enzymatic hydrolysis Cell Line Tumor medicine Animals Humans Serum Albumin Fluorescent Dyes Molecular Structure 010405 organic chemistry Chemistry Organic Chemistry Optical Imaging General Chemistry Neoplasms Experimental 0104 chemical sciences Kinetics medicine.anatomical_structure Membrane protein Biophysics Ex vivo Intracellular Protein ligand |
| Zdroj: | Chemistry, an Asian journal. 16(8) |
| ISSN: | 1861-471X |
| Popis: | To date, various affinity-based protein labeling probes have been developed and applied in biological research to modify endogenous proteins in cell lysates and on the cell surface. However, the reactive groups on the labeling probes are also the cause of probe instability and nonselective labeling in a more complex environment, e. g., intracellular and in vivo. Here, we show that labeling probes composed of a sterically stabilized difluorophenyl pivalate can achieve efficient and selective labeling of endogenous proteins on the cell surface, inside living cells and in vivo. As compared with the existing protein labeling probes, probes with the difluorophenyl pivalate exhibit several advantages, including long-term stability in stock solutions, resistance to enzymatic hydrolysis and can be customized easily with diverse fluorophores and protein ligands. With this probe design, endogenous hypoxia biomarker in living cells and nude mice were successfully labeled and validated by in vivo, ex vivo, and immunohistochemistry imaging. |
| Databáze: | OpenAIRE |
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