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A new GFP fluorophore-based probe for lysosomes labelling and tracing lysosomal viscosity in live cells

Abstract

Lysosomes, the main digestive compartment of the live cells, are the most acidic organelles (pH 3.8-6.6) with the highest viscosity (47-190 cP at 25 ℃). Lysosomal viscosity can reflect the status and function of the lysosomes, and the abnormal of lysosomal viscosity is responsible for the dysfunction of lysosomes, which is closely related to multiple diseases. In this work, a new GFP (green fluorescent protein) fluorophore-based probe (Lys-V) was designed and synthesized for mapping lysosomal viscosity in live cells. The increasing viscosity can restrict the C=C twist around the exocyclic double bond between phenol and imidazolidinone, so the fluorescence emission of Lys-V would enhance with the increasing of viscosity, and there is an excellent linear relationship between the Log FI (515 nm) and the Log Viscosity. Notably, fluorescence of Lys-V is quite stable in different pH solvents at same viscosi3ty, demonstrating that Lys-V is suitable for quantifying viscosity in a wide pH range (from 4 to 10). Because of its ideal water solubility, low cytotoxicity and weak basic amines morpholine substituents, Lys-V can be utilized for lysosome labelling in live cells. Significantly, the dynamic viscosity changes in lysosomes under dexamethasone stimulus could also be captured by Lys-V. Our results will allow Lys-V to become a new chemical tool for detecting lysosomal viscosity in live cells as well as unveiling the underlying mechanism of lysosome behaviour during various biological processes, which is beneficial for studying lysosome-related diseases as well as exploration of lysosome-related pharmacology.

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Publication details

The article was received on 20 Jul 2018, accepted on 10 Sep 2018 and first published on 12 Sep 2018


Article type: Paper
DOI: 10.1039/C8TB01885E
Citation: J. Mater. Chem. B, 2018, Accepted Manuscript
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    A new GFP fluorophore-based probe for lysosomes labelling and tracing lysosomal viscosity in live cells

    X. Li, R. Zhao, Y. Wang and C. Huang, J. Mater. Chem. B, 2018, Accepted Manuscript , DOI: 10.1039/C8TB01885E

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