Discrimination of cellular developmental states focusing on glycan transformation and membrane dynamics by using BODIPY-tagged lactosyl ceramides.
Kenta AraiAtsuko OhtakeShusaku DaikokuKatsuhiko SuzukiYukishige ItoKazuya KabayamaKoichi FukaseYoshimi KanieOsamu KaniePublished in: Organic & biomolecular chemistry (2021)
Glycosphingolipids (GSLs) are a group of molecules composed of a hydrophilic glycan part and a hydrophobic ceramide creating a diverse family. GSLs are de novo synthesised from ceramides at the endoplasmic reticulum and Golgi apparatus, and transported to the outer surface of the plasma membrane. It has been known that the glycan structures of GSLs change reflecting disease states. We envisioned that analysing the glycan pattern of GSLs enables distinguishing diseases. For this purpose, we utilised a fluorescently tagged compound, LacCerBODIPY (1). At first, compound 1 was taken up by cultured PC12D cells and transformed into various GSLs. As a result, changes in the GSL patterns of differentiation states of the cells were successfully observed by using an analysis platform, nano-liquid chromatography (LC)-fluorescence detection (FLD)-electrospray ionisation (ESI)-mass spectrometry (MS), which could quantify and provide molecular ions simultaneously. We found that compound 1 remained for about 10 min on the plasma membrane before it was converted into other GSLs. We therefore investigated a more rapid way to discriminate different cellular states by fluorescence recovery after photobleaching, which revealed that it is possible to distinguish the differentiation states as well.
Keyphrases
- mass spectrometry
- liquid chromatography
- endoplasmic reticulum
- high resolution mass spectrometry
- induced apoptosis
- tandem mass spectrometry
- simultaneous determination
- high resolution
- ms ms
- cell cycle arrest
- gas chromatography
- cell surface
- single molecule
- high performance liquid chromatography
- capillary electrophoresis
- loop mediated isothermal amplification
- solid phase extraction
- single cell
- high throughput
- endothelial cells
- signaling pathway
- ionic liquid
- energy transfer
- cell death
- atomic force microscopy
- aqueous solution