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Heterocyclic sterol probes for live monitoring of sterol trafficking and lysosomal storage disorders
J. Králová, M. Jurášek, L. Krčová, B. Dolenský, I. Novotný, M. Dušek, Z. Rottnerová, M. Kahle, P. Drašar, P. Bartůněk, V. Král,
Language English Country Great Britain
Document type Journal Article, Research Support, Non-U.S. Gov't
Grant support
17-02836S
Grantová Agentura České Republiky (Grant Agency of the Czech Republic) - International
LO1220
Ministerstvo Školství, Mládeže a Tělovýchovy (Ministry of Education, Youth and Sports) - International
NLK
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- MeSH
- Biological Transport MeSH
- Cell Membrane chemistry metabolism MeSH
- Cell Line MeSH
- Cholesterol analysis metabolism MeSH
- Fluorescent Dyes chemistry MeSH
- Microscopy, Fluorescence methods MeSH
- Humans MeSH
- Lysosomal Storage Diseases diagnosis metabolism MeSH
- Pyridines chemistry MeSH
- Boron Compounds chemistry MeSH
- Sterols chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The monitoring of intracellular cholesterol homeostasis and trafficking is of great importance because their imbalance leads to many pathologies. Reliable tools for cholesterol detection are in demand. This study presents the design and synthesis of fluorescent probes for cholesterol recognition and demonstrates their selectivity by a variety of methods. The construction of dedicated library of 14 probes was based on heterocyclic (pyridine)-sterol derivatives with various attached fluorophores. The most promising probe, a P1-BODIPY conjugate FP-5, was analysed in detail and showed an intensive labelling of cellular membranes followed by intracellular redistribution into various cholesterol rich organelles and vesicles. FP-5 displayed a stronger signal, with faster kinetics, than the commercial TF-Chol probe. In addition, cells with pharmacologically disrupted cholesterol transport, or with a genetic mutation of cholesterol transporting protein NPC1, exhibited strong and fast FP-5 signal in the endo/lysosomal compartment, co-localizing with filipin staining of cholesterol. Hence, FP-5 has high potential as a new probe for monitoring cholesterol trafficking and its disorders.
References provided by Crossref.org
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