Green Fluorescent Protein GFP-Chromophore-Based Probe for the Detection of Mitochondrial Viscosity in Living Cells

Abstract

Viscosity is a pivotal factor for indicating the dysfunction of the mitochondria. To date, most of the fluorescent probes developed for mitochondrial viscosity have been designed using BODIPY, hemicyanine, or pyridine-based molecular rotors as part of the core structure. Our aim with this research was to extend the range of suitable fluorophores available for the construction of such fluorescent molecular rotors for evaluating the viscosity of mitocondria. Herein, we have developed a green fluorescent protein (GFP)-chromophore-based fluorescent probe (MIT-V) for the detection of mitochondrial viscosity in live cells. MIT-V exhibited a high sensitivity toward viscosity (from 7.9 cP to 438.4 cP). The “off–on” sensing mechanism of MIT-V was ascribed to the restricted rotation of single bonds and excited-state CC double bonds of MIT-V. Cell studies indicated that MIT-V targets the mitochondria and that it was able to monitor real-time changes in the viscosity of live HeLa cell mitochondria. Therefore, we propose that MIT-V can be used as an effective chemosensor for the real-time imaging of mitochondrial viscosity in live cells. Our results clearly demonstrate the utility of such GFP-chromophore-based derivatives for the development of viscosity-sensitive systems.

Super-resolution analyzing spatial organization of lysosomes with an organic fluorescent probe

“…[8] Especially, the viscosity of mitochondria and lysosomes in live cells has been reported. [9] However, few studies have been dedicated to reveal the spatial organization between lysosomes and mitochondria using SIM techniques.…”

A Fluorogenic Far Red-Emitting Molecular Viscometer for Ascertaining Lysosomal Stress in Live Cells and Caenorhabditis elegans

“…It is an acidic compartment and contains approximately 60 hydrolytic enzymes for breaking all sorts of biomolecules ( Lawrence and Zoncu, 2019 ). The viscosity of lysosome and lysosome-related organelles (LROs) is closely associated with overall animal health and a key indicator of its functionality ( Li et al, 2018 ; Tan et al, 2019 ; Cai et al, 2021 ). Therefore, sensitive monitoring of their viscosity with a specific nano-sized molecular viscometer is essential for understanding cellular health.…”

Fabrication of multifunctional metal–organic frameworks nanoparticles via layer-by-layer self-assembly to efficiently discover PSD95-nNOS uncouplers for stroke treatment

“…Considering their complicated operations and the expensive cost [ 15 , 33 , 34 ], another rapid and sensitive method need be constructed. In recent years, fluorescent tags such as green fluorescent protein (GFP), red fluorescent protein (RFP) and yellow fluorescent protein (YFP) are commonly used in protein optical detection to enhance the detection sensitivity [ 35 , 36 ], furthermore, fluorescence-based technologies have shown great characteristics of high sensitivity, rapid response and easy operation for high-throughput screening [ 37 ], therefore, it is a good choice to apply fluorescent tags to label PSD95 for more efficient detection.…”