Citation: Xiangning Fang, Qinglong Qiao, Fei Deng, Zhaochao Xu. Designing malachite green derivatives to optimize fluorogen-activating protein pairs for rapid PAINT super-resolution imaging in living cells[J]. Chinese Chemical Letters, ;2025, 36(12): 110980. doi: 10.1016/j.cclet.2025.110980 shu

Designing malachite green derivatives to optimize fluorogen-activating protein pairs for rapid PAINT super-resolution imaging in living cells

Xiangning Fang ^{a, b} ,
Qinglong Qiao ^{a, *,} ,
Fei Deng ^{c, d} ,
Zhaochao Xu ^{a, b, *,}

a.
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
b.
University of Chinese Academy of Sciences, Beijing 100049, China
c.
School of Chemistry and Chemical Engineering, Jinggangshan University, Ji'an 343009, China
d.
Key Laboratory of Jiangxi Province for Special Optoelectronic Artificial Crystal Materials, Jinggangshan University, Ji'an 343009, China

qqlqiao@dicp.ac.cn

zcxu@dicp.ac.cn

Received Date: 11 January 2025
Revised Date: 18 February 2025
Accepted Date: 19 February 2025
Available Online: 19 February 2025

Figures(4)

Fluorogen-activating proteins (FAPs) selectively bind to specific fluorophores, inducing fluorescence activation through the inhibition of torsion of fluorophores. This binding-activation mechanism provides a highly specific and efficient fluorescence system that minimizes background signals, significantly enhancing the signal-to-noise ratio (SNR) and making it a powerful tool in live-cell imaging. The principle of binding-activation fluorescence is fundamental to point accumulation for imaging in nanoscale topography (PAINT) super-resolution imaging. However, the high binding affinity between traditional FAP-fluorophore pairs limits their application in PAINT, thus hindering the rapid and dynamic imaging necessary for high-resolution cellular studies. In this work, we designed malachite green (MG) derivatives with bulky N-substituents to modulate the binding affinity of the MG-dL5** fluorophore-FAP pair. This modification introduces steric hindrance in MG-dL5** system, resulting in reduced binding affinity and practicability for fast, high-resolution PAINT imaging. Among the synthesized derivatives, MG-Pen showed optimal properties, enabling rapid and high-resolution PAINT imaging of dL5** in living cells. This study highlights the potential of MG derivatives optimization in overcoming the limitations of fluorophore-FAP pairs for super-resolution imaging and provides a new approach for enhancing the performance of PAINT in living cell applications.
- Malachite green,
- dL5**,
- PAINT,
- Super-resolution imaging,
- Fluorogen-activating protein
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