Synthesis and biological properties of the fluorescent ether lipid precursor 1-O-[9′-(1″-pyrenyl)]nonyl-sn-glycerol

Journal of Lipid Research

Richard I. Duclos, Jr.¹  Harrison W. Farber¹  Conor C. Smith²  Raphael A. Zoeller²  Hongying Zheng² 

[1] Pulmonary Center, Boston University School of Medicine, Boston, MA 02118-2526;Department of Physiology and Biophysics, Boston University School of Medicine, Boston, MA 02118-2526;

关键词: pyrene;    fluorescence;    chemical synthesis;    ether lipid;    plasmalogen;    alkylglycerol;   

DOI  :  

来源: DOAJ

【 摘 要 】

The synthesis of an ω-pyrene-labeled 1-O-alkyl-sn-glycerol was performed using a chirospecific method starting from R-(−)-2,3-O-isopropylidene-sn-glycerol. The product, 1-O-[9′-(1″-pyrenyl)]nonyl-sn-glycerol (pAG), is a fluorescent ether lipid that has a pyrene moiety covalently attached at the alkyl chain terminus. pAG was taken into CHO-K1 cells and a plasmalogen-deficient variant of CHO-K1, NRel-4. This variant is defective in dihydroxyacetonephosphate acyltransferase, which catalyzes the first step in plasmenylethanolamine (PlsEtn) biosynthesis. pAG was incorporated primarily into ethanolamine and choline phospholipids as well as a neutral lipid fraction tentatively identified as alkyldiacylglycerol. NRel-4 accumulated more fluorescence in the phospholipid fraction than CHO-K1, specifically in the ethanolamine phospholipids. Analysis of the fluorescent lipids showed that 93% of the pAG was incorporated into glycerolipids with the ether bond intact. Although the addition of 20 μM 1-O-hexadecyl-sn-glycerol to the medium fully restored PlsEtn biosynthesis in NRel-4 cells, pAG only partially restored PlsEtn synthesis. Incubation of cells with pAG followed by irradiation with long-wavelength (>300 nm) ultraviolet light resulted in cytotoxicity. NRel-4 cells displayed an increased sensitivity to this treatment compared with CHO-K1 cells. This photodynamic cytotoxicity approach could be used to select for mutants that are defective in downstream steps in ether lipid biosynthesis.

【 授权许可】

Unknown