Vacuole-Specific Lipid Release for Tracking Intracellular Lipid Metabolism and Transport in Saccharomyces cerevisiae .
Vladimir GirikSuihan FengHanaa HaririW Mike HenneHoward RiezmanPublished in: ACS chemical biology (2022)
Lipid metabolism is spatiotemporally regulated within cells, yet intervention into lipid functions at subcellular resolution remains difficult. Here, we report a method that enables site-specific release of sphingolipids and cholesterol inside the vacuole in Saccharomyces cerevisiae . Using this approach, we monitored real-time sphingolipid metabolic flux out of the vacuole by mass spectrometry and found that the endoplasmic reticulum-vacuole-tethering protein Mdm1 facilitated the metabolism of sphingoid bases into ceramides. In addition, we showed that cholesterol, once delivered into yeast using our method, could restore cell proliferation induced by ergosterol deprivation, overcoming the previously described sterol-uptake barrier under aerobic conditions. Together, these data define a new way to study intracellular lipid metabolism and transport from the vacuole in yeast.
Keyphrases
- electronic health record
- saccharomyces cerevisiae
- fatty acid
- endoplasmic reticulum
- mass spectrometry
- cell proliferation
- randomized controlled trial
- induced apoptosis
- low density lipoprotein
- cell cycle arrest
- transcription factor
- high resolution
- pi k akt
- oxidative stress
- deep learning
- endoplasmic reticulum stress
- cell death
- high performance liquid chromatography
- ms ms
- big data