Tracking the Subcellular Fate of 20(S)-Hydroxycholesterol with Click Chemistry Reveals a Transport Pathway to the Golgi

Peyrot, Sara M.; Nachtergaele, Sigrid; Luchetti, Giovanni; Mydock-McGrane, Laurel; Fujiwara, Hideji; Scherrer, David E.; Jallouk, Andrew P.; Schlesinger, Paul H.; Ory, Daniel S.; Covey, Douglas F.; Rohatgi, Rajat

doi:10.1074/jbc.m113.540351

Cited by 22 publications

(30 citation statements)

References 73 publications

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“…The difference in the cell. Thus, NPC1 is required for effi cient traffi cking of 25-HCTL from lysosomes, similar to what was previously reported for an alkyne oxysterol analog ( 40 ).…”

Section: -Hc Has a Distribution Similar To Cholesterolsupporting

confidence: 57%

“…This distribution is similar to that of the fl uorescent cholesterol probes, DHE ( 32 ) and CTL ( 46 ), yet markedly different from the previously reported 20(S)-hydroxycholesterol alkyne probe ( 40 ). This fi nding suggests that 25-HCTL, like other sterol probes ( 40,44 ), initially transits through the lysosome and then redistributes among membranes (e.g., ERC and plasma membrane) to achieve a steady-state distribution similar to that of cholesterol. 25-HCTL is also esterifi ed and incorporated into lipid droplets, indicating that, like 25-HC, the analog traffi cs to the ER.…”

Section: Discussionmentioning

confidence: 52%

“…This is supported by experiments demonstrating that 25-HCTL partitions into serum lipoproteins or purifi ed LDL and enters the cell via LDLR, rather than through passive diffusion. This mechanism may be shared with other oxysterol probes that also initially transit through the lysosome and then redistribute to other organelles ( 40,44 ).…”

Section: Discussionmentioning

confidence: 99%

“…approaches have been successfully used. One approach is the introduction of an alkyne moiety to lipids that can be conjugated to a fl uorophore using bio-orthognal chemistry ( 40,44,45 ). Although the alkyne moiety does not greatly alter the biophysical properties of the sterols, it precludes live cell imaging due to the conjugation requirements ( 40,44 ).…”

Section: -Hc Has a Distribution Similar To Cholesterolmentioning

confidence: 99%

“…One approach is the introduction of an alkyne moiety to lipids that can be conjugated to a fl uorophore using bio-orthognal chemistry ( 40,44,45 ). Although the alkyne moiety does not greatly alter the biophysical properties of the sterols, it precludes live cell imaging due to the conjugation requirements ( 40,44 ). An alternative approach takes advantage of intrinsically fl uorescent sterols, including the naturally occurring cholesterol analog, dehydroergosterol (DHE) ( 32 ), or a synthetic sterol, CTL ( 46 ).…”

Section: -Hc Has a Distribution Similar To Cholesterolmentioning

confidence: 99%

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A novel intrinsically fluorescent probe for study of uptake and trafficking of 25-hydroxycholesterol

Iaea

Gale

Bielska

et al. 2015

Journal of Lipid Research

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This article is available online at http://www.jlr.org by coordinated homeostatic mechanisms, primarily involving the regulated proteolysis of the sterol regulatory element-binding protein (SREBP)-2 transcription factor in the Golgi ( 1 ). When ER sterols are abundant, SREBP-2 is retained in the ER in a complex with the cholesterol-sensing protein, SREBP cleavage-activating protein (SCAP), and the ER retention proteins, Insig-1 or -2 ( 2, 3 ). When cholesterol content in the ER is depleted, SCAP undergoes a conformational change and is released from Insig, allowing the SREBP-SCAP complex to translocate to the Golgi apparatus ( 4, 5 ) where it undergoes proteolytic maturation with release of the SREBP transcription factor ( 6 ).Cholesterol homeostasis is regulated not only by cholesterol, but also by oxygenated cholesterol species, referred to as oxysterols ( 1, 7-9 ) Side-chain oxysterols, such as 25-hydroxycholesterol (25-HC), are generated enzymatically and act as important regulators of cholesterol homeostasis, despite their presence in cells being only at 0.1% the concentration of cholesterol ( 9 ). At the transcriptional level, 25-HC can bind the liver X receptors (LXRs) to active LXR-mediated transcription that results in increased cholesterol effl ux and elimination ( 10, 11 ). 25-HC also inhibits SREBP maturation and subsequent transcription of genes involved in cholesterol biosynthesis and uptake ( 1 ). In contrast to cholesterol, which directly interacts with SCAP, 25-HC enhances the interaction between SCAP and Insig proteins, resulting in retention of the SREBP-2-SCAP complex in the ER ( 12, 13 ).

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“…The difference in the cell. Thus, NPC1 is required for effi cient traffi cking of 25-HCTL from lysosomes, similar to what was previously reported for an alkyne oxysterol analog ( 40 ).…”

Section: -Hc Has a Distribution Similar To Cholesterolsupporting

confidence: 57%