Citation: Yefeng Chen, Chenghong Xue, Jie Wang, Minqiu Xu, Yuyao Li, Yiru Ding, Heng Song, Weipan Xu, Hexin Xie. High-contrast and real-time visualization of membrane proteins in live cells with malachite green-based fluorogenic probes[J]. Chinese Chemical Letters, ;2022, 33(3): 1637-1642. doi: 10.1016/j.cclet.2021.09.088 shu

High-contrast and real-time visualization of membrane proteins in live cells with malachite green-based fluorogenic probes

State Key Laboratory of Bioreactor Engineering, Shanghai Key Laboratory of New Drug Design, Frontiers Science Center for Materiobiology and Dynamic Chemistry, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China

xiehexin@ecust.edu.cn

Received Date: 8 July 2021
Revised Date: 19 September 2021
Accepted Date: 24 September 2021
Available Online: 30 September 2021

Figures(6)

Imaging dynamics of membrane proteins of live cells in a wash-free and real-time manner has been a challenging task. Herein, we report unprecedented applications of malachite green (MG), an organic dye widely used in pigment industry, as a switchable fluorophore to monitor membrane enzymes or non-catalytic proteins in live cells. Conformationally flexible MG is non-fluorescent in aqueous solution, yet covalent binding with endogenous proteins of cells significantly enhances its fluorescence at 670 nm by restricting flexibility of dye. Integrating a phosphate-caged quinone methide precursor with MG yielded a covalent labeling fluorogenic probe, allowing real-time imaging of membrane alkaline phosphatase (ALP, a model catalytic protein) activity in live cells with over 100-fold enhancement of fluorescence intensity. Moreover, MG is also applicable to image non-catalytic protein by conjugation with protein-specific ligand. A fluorogenic probe consisted of c-RGDfK peptide and MG proved to be compatible with wash-free and real-time visualization of non-catalytic integrin α_vβ₃ in live cells with high contrast.
- Cellular imaging,
- Fluorogenic probe,
- Environment-sensitive fluorophore,
- Alkaline phosphatase,
- Integrin
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