A novel alkyne cholesterol to trace cellular cholesterol metabolism and localization
| Autor: | Yuriy Kiver, Lars Kuerschner, Christoph Thiele, Mario Schoene, Anne Gaebler, Silvia Friedrichs, Dieter Lütjohann, Hans-Frieder Schött, Kristina Hofmann |
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| Rok vydání: | 2014 |
| Předmět: |
Cholesterol oxidase
Golgi Apparatus Endoplasmic Reticulum Cholesterol analog Biochemistry Cell membrane chemistry.chemical_compound Endocrinology Methods alkyne lipid analog Molecular Structure Mitochondria Cell biology Sterols Cholesterol medicine.anatomical_structure Cell Tracking Alkynes Click chemistry symbols oxysterols lipids (amino acids peptides and proteins) Cholesterol Esters click reaction 5-Aminolevulinate Synthetase Saccharomyces cerevisiae Proteins Saccharomyces cerevisiae probe QD415-436 Biology symbols.namesake Cell Line Tumor medicine Animals Humans Cholesterol Oxidase Endoplasmic reticulum Cell Membrane Cell Biology Metabolism Golgi apparatus Rats Kinetics Microscopy Fluorescence chemistry Mutation Acyltransferases |
| Zdroj: | Journal of Lipid Research, Vol 55, Iss 3, Pp 583-591 (2014) |
| ISSN: | 0022-2275 |
| DOI: | 10.1194/jlr.d044727 |
| Popis: | Cholesterol is an important lipid of mammalian cells and plays a fundamental role in many biological processes. Its concentration in the various cellular membranes differs and is tightly regulated. Here, we present a novel alkyne cholesterol analog suitable for tracing both cholesterol metabolism and localization. This probe can be detected by click chemistry employing various reporter azides. Alkyne cholesterol is accepted by cellular enzymes from different biological species (Brevibacterium, yeast, rat, human) and these enzymes include cholesterol oxidases, hydroxylases, and acyl transferases that generate the expected metabolites in in vitro and in vivo assays. Using fluorescence microscopy, we studied the distribution of cholesterol at subcellular resolution, detecting the lipid in the Golgi and at the plasma membrane, but also in the endoplasmic reticulum and mitochondria. In summary, alkyne cholesterol represents a versatile, sensitive, and easy-to-use tool for tracking cellular cholesterol metabolism and localization as it allows for manifold detection methods including mass spectrometry, thin-layer chromatography/fluorography, and fluorescence microscopy. |
| Databáze: | OpenAIRE |
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