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

Lei Cai; Huan Li; Xiang Yu; Luling Wu; Xiaoqin Wei; Tony D. James; Chusen Huang

doi:10.1021/acsabm.0c01446

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

Lei Cai, Huan Li, Xiang Yu, Luling Wu, Xiaoqin Wei, Tony D. James, Chusen Huang

Department of Chemistry

Research output: Contribution to journal › Article › peer-review

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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.

Original language	English
Pages (from-to)	2128-2134
Number of pages	7
Journal	ACS Applied Bio Materials
Volume	4
Issue number	3
Early online date	14 Jan 2021
DOIs	https://doi.org/10.1021/acsabm.0c01446
Publication status	Published - 15 Mar 2021

Bibliographical note

Funding Information:
C.H. thanks the National Natural Science Foundation of China (Grants 21672150, 21302125), Alexander von Humboldt Foundation (AvH), Shanghai Rising-Star Program (19QA1406400), Shanghai Government (18DZ2254200), Program for Changjiang Scholars and Innovative (IRT_16R49). T.D.J. wishes to thank the Royal Society for a Wolfson Research Merit Award and the Open Research Fund of the School of Chemistry and Chemical Engineering, Henan Normal University for support (2020ZD01). L.W. wishes to thank the China Scholarship Council and the University of Bath for supporting his PhD in the United Kingdom.

Keywords

fluorescent probe
GFP-chromophore
live cell imaging
mitochondria
viscosity

ASJC Scopus subject areas

Biomaterials
General Chemistry
Biomedical Engineering
Biochemistry, medical

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10.1021/acsabm.0c01446

GFP Submitted December 2020.PDF
This document is the Accepted Manuscript version of a Published Work that appeared in final form in Applied Bio Materials, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acsabm.0c01446
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Cite this

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title = "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.",

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author = "Lei Cai and Huan Li and Xiang Yu and Luling Wu and Xiaoqin Wei and James, {Tony D.} and Chusen Huang",

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AU - Wei, Xiaoqin

AU - James, Tony D.

AU - Huang, Chusen

N1 - Funding Information: C.H. thanks the National Natural Science Foundation of China (Grants 21672150, 21302125), Alexander von Humboldt Foundation (AvH), Shanghai Rising-Star Program (19QA1406400), Shanghai Government (18DZ2254200), Program for Changjiang Scholars and Innovative (IRT_16R49). T.D.J. wishes to thank the Royal Society for a Wolfson Research Merit Award and the Open Research Fund of the School of Chemistry and Chemical Engineering, Henan Normal University for support (2020ZD01). L.W. wishes to thank the China Scholarship Council and the University of Bath for supporting his PhD in the United Kingdom.

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

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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