Influence of fluorophore and linker length on the localization and trafficking of fluorescent sterol probes

Abstract Fluorescent sterol probes, comprising a fluorophore connected to a sterol backbone by means of a linker, are promising tools for enabling high-resolution imaging of intracellular cholesterol. In this study, we evaluated how the size of the linker, site of its attachment and nature of the fl...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Jarmila Králová, Michal Jurášek, Lucie Mikšátková, Anna Marešová, Jan Fähnrich, Petra Cihlářová, Pavel Drašar, Petr Bartůněk, Vladimír Král
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9f4dc5143084469ea4450e9de614eba2
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:9f4dc5143084469ea4450e9de614eba2
record_format	dspace
spelling	oai:doaj.org-article:9f4dc5143084469ea4450e9de614eba22021-12-02T11:57:56ZInfluence of fluorophore and linker length on the localization and trafficking of fluorescent sterol probes10.1038/s41598-020-78085-92045-2322https://doaj.org/article/9f4dc5143084469ea4450e9de614eba22020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78085-9https://doaj.org/toc/2045-2322Abstract Fluorescent sterol probes, comprising a fluorophore connected to a sterol backbone by means of a linker, are promising tools for enabling high-resolution imaging of intracellular cholesterol. In this study, we evaluated how the size of the linker, site of its attachment and nature of the fluorophore, affect the localization and trafficking properties of fluorescent sterol probes. Varying lengths of linker using the same fluorophore affected cell penetration and retention in specific cell compartments. A C-4 linker was confirmed as optimal. Derivatives of heterocyclic sterol precursors attached with identical C-4 linker to different fluorophores at diverse positions also showed significant differences in their binding properties to various intracellular compartments and kinetics of trafficking. Two novel red-emitting probes with good cell permeability, fast intracellular labelling and slightly different distribution displayed very promising characteristics for sterol probes. These probes also strongly labelled endo/lysosomal compartment in cells with pharmacologically disrupted cholesterol transport, or with a genetic mutation of cholesterol transporting protein NPC1, that overlapped with filipin staining of cholesterol. Overall, the present study demonstrates that the physicochemical properties of the fluorophore/linker pairing determine the kinetics of uptake and distribution and subsequently influence the applicability of final probes.Jarmila KrálováMichal JurášekLucie MikšátkováAnna MarešováJan FähnrichPetra CihlářováPavel DrašarPetr BartůněkVladimír KrálNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-11 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jarmila Králová Michal Jurášek Lucie Mikšátková Anna Marešová Jan Fähnrich Petra Cihlářová Pavel Drašar Petr Bartůněk Vladimír Král Influence of fluorophore and linker length on the localization and trafficking of fluorescent sterol probes
description	Abstract Fluorescent sterol probes, comprising a fluorophore connected to a sterol backbone by means of a linker, are promising tools for enabling high-resolution imaging of intracellular cholesterol. In this study, we evaluated how the size of the linker, site of its attachment and nature of the fluorophore, affect the localization and trafficking properties of fluorescent sterol probes. Varying lengths of linker using the same fluorophore affected cell penetration and retention in specific cell compartments. A C-4 linker was confirmed as optimal. Derivatives of heterocyclic sterol precursors attached with identical C-4 linker to different fluorophores at diverse positions also showed significant differences in their binding properties to various intracellular compartments and kinetics of trafficking. Two novel red-emitting probes with good cell permeability, fast intracellular labelling and slightly different distribution displayed very promising characteristics for sterol probes. These probes also strongly labelled endo/lysosomal compartment in cells with pharmacologically disrupted cholesterol transport, or with a genetic mutation of cholesterol transporting protein NPC1, that overlapped with filipin staining of cholesterol. Overall, the present study demonstrates that the physicochemical properties of the fluorophore/linker pairing determine the kinetics of uptake and distribution and subsequently influence the applicability of final probes.
format	article
author	Jarmila Králová Michal Jurášek Lucie Mikšátková Anna Marešová Jan Fähnrich Petra Cihlářová Pavel Drašar Petr Bartůněk Vladimír Král
author_facet	Jarmila Králová Michal Jurášek Lucie Mikšátková Anna Marešová Jan Fähnrich Petra Cihlářová Pavel Drašar Petr Bartůněk Vladimír Král
author_sort	Jarmila Králová
title	Influence of fluorophore and linker length on the localization and trafficking of fluorescent sterol probes
title_short	Influence of fluorophore and linker length on the localization and trafficking of fluorescent sterol probes
title_full	Influence of fluorophore and linker length on the localization and trafficking of fluorescent sterol probes
title_fullStr	Influence of fluorophore and linker length on the localization and trafficking of fluorescent sterol probes
title_full_unstemmed	Influence of fluorophore and linker length on the localization and trafficking of fluorescent sterol probes
title_sort	influence of fluorophore and linker length on the localization and trafficking of fluorescent sterol probes
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/9f4dc5143084469ea4450e9de614eba2
work_keys_str_mv	AT jarmilakralova influenceoffluorophoreandlinkerlengthonthelocalizationandtraffickingoffluorescentsterolprobes AT michaljurasek influenceoffluorophoreandlinkerlengthonthelocalizationandtraffickingoffluorescentsterolprobes AT luciemiksatkova influenceoffluorophoreandlinkerlengthonthelocalizationandtraffickingoffluorescentsterolprobes AT annamaresova influenceoffluorophoreandlinkerlengthonthelocalizationandtraffickingoffluorescentsterolprobes AT janfahnrich influenceoffluorophoreandlinkerlengthonthelocalizationandtraffickingoffluorescentsterolprobes AT petracihlarova influenceoffluorophoreandlinkerlengthonthelocalizationandtraffickingoffluorescentsterolprobes AT paveldrasar influenceoffluorophoreandlinkerlengthonthelocalizationandtraffickingoffluorescentsterolprobes AT petrbartunek influenceoffluorophoreandlinkerlengthonthelocalizationandtraffickingoffluorescentsterolprobes AT vladimirkral influenceoffluorophoreandlinkerlengthonthelocalizationandtraffickingoffluorescentsterolprobes
_version_	1718394824460075008

Influence of fluorophore and linker length on the localization and trafficking of fluorescent sterol probes

Ejemplares similares