title: A novel alkyne cholesterol to trace cellular cholesterol metabolism and localization.
abstract: 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.
authors: Life and Medical Sciences Institute (LIMES), University of Bonn, D-53115 Bonn, Germany; and.
authors: Kristina
authors: K
authors: Hofmann
authors: Christoph
authors: C
authors: Thiele
authors: Hans-Frieder
authors: HF
authors: Schött
authors: Anne
authors: A
authors: Gaebler
authors: Mario
authors: M
authors: Schoene
authors: Yuriy
authors: Y
authors: Kiver
authors: Silvia
authors: S
authors: Friedrichs
authors: Dieter
authors: D
authors: Lütjohann
authors: Lars
authors: L
authors: Kuerschner
mesh_terms: D000624:5-Aminolevulinate Synthetase / Q000235:genetics; D000217:Acyltransferases / Q000378:metabolism; D000480:Alkynes / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D058948:Cell Tracking / Q000379:methods*; D002784:Cholesterol / Q000031:analogs & derivatives / Q000737:chemistry* / Q000378:metabolism*; D002788:Cholesterol Esters / Q000378:metabolism; D002789:Cholesterol Oxidase / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism; D015394:Molecular Structure; D009154:Mutation; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D013261:Sterols / Q000378:metabolism
chemical_list: C000592552:25-ethinyl-26-nor-3-hydroxycholest-5-ene; D000480:Alkynes; D002788:Cholesterol Esters; D029701:Saccharomyces cerevisiae Proteins; D013261:Sterols; D002784:Cholesterol; D002789:Cholesterol Oxidase; D000217:Acyltransferases; D000624:5-Aminolevulinate Synthetase; C577889:aminolevulinate synthase, S cerevisiae
keywords: alkyne lipid; analog; click reaction; oxysterols; probe