title: Synthesis and biological properties of the fluorescent ether lipid precursor 1-O-[9'-(1''-pyrenyl)]nonyl-sn-glycerol.
abstract: The synthesis of an omega-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 microM 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.
authors.affiliation: Department of Physiology and Biophysics, Boston University School of Medicine, Boston, MA 02118-2526, USA.
authors.forename: Hongying
authors.initials: H
authors.lastname: Zheng
authors.forename: Richard I
authors.initials: RI
authors.lastname: Duclos
authors.forename: Conor C
authors.initials: CC
authors.lastname: Smith
authors.forename: Harrison W
authors.initials: HW
authors.lastname: Farber
authors.forename: Raphael A
authors.initials: RA
authors.lastname: Zoeller
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D005453:Fluorescence; D005990:Glycerol / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology; D010742:Phospholipid Ethers / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D010743:Phospholipids / Q000096:biosynthesis; D011721:Pyrenes / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D014466:Ultraviolet Rays
chemical_list: C512135:1-O-(9'-(1''-pyrenyl))nonylglycerol; D010742:Phospholipid Ethers; D010743:Phospholipids; D011721:Pyrenes; D005990:Glycerol