title: Fluorescent polyene ceramide analogues as membrane probes.
abstract: Three ceramide analogues have been synthesized, with sphingosine-like chains containing five conjugated double bonds. Pentaene I has an N-palmitoyl acyl chain, while the other two pentaenes contain also a doxyl radical, respectively, at C5 (Penta5dox) and at C16 (Penta16dox) positions of the N-acyl chain. Pentaene I maximum excitation and emission wavelengths in a phospholipid bilayer are 353 and 478 nm, respectively. Pentaene I does not segregate from the other lipids in the way natural ceramide does, but rather mixes with them in a selective way according to the lipid phases involved. Fluorescence confocal microscopy studies show that when lipid domains in different physical states coexist, Pentaene I emission is higher in gel than in fluid domains, and in liquid-ordered than in liquid-disordered areas. Electron paramagnetic resonance of the pentaene doxyl probes confirms that these molecules are sensitive to the physical state of the bilayer. Calorimetric and fluorescence quenching experiments suggest that the lipids under study orient themselves in lipid bilayers with their polar moieties located at the lipid-water interface. The doxyl radical in the N-acyl chain quenches the fluorescence of the pentaene group when in close proximity. Because of this property, Penta16dox can detect gel-fluid transitions in phospholipids. The availability of probes for lipids in the gel phase is important in view of novel evidence for the existence of gel microdomains in cell membranes.
authors: Spanish National Research Council (CSIC), Institute of Advanced Chemistry of Catalonia (IQACCSIC), Department of Biomedicinal Chemistry, Research Unit on Bioactive Molecules (RUBAM), Jordi Girona 1826, 08034 Barcelona, Spain.
authors: Ingrid
authors: I
authors: Nieves
authors: Ibai
authors: I
authors: Artetxe
authors: José Luis
authors: JL
authors: Abad
authors: Alicia
authors: A
authors: Alonso
authors: Jon V
authors: JV
authors: Busto
authors: Lluís
authors: L
authors: Fajarí
authors: L Ruth
authors: LR
authors: Montes
authors: Jesús
authors: J
authors: Sot
authors: Antonio
authors: A
authors: Delgado
authors: Félix M
authors: FM
authors: Goñi
mesh_terms: D002518:Ceramides / Q000138:chemical synthesis / Q000737:chemistry*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D015394:Molecular Structure; D011090:Polyenes / Q000138:chemical synthesis / Q000737:chemistry*
chemical_list: D002518:Ceramides; D005456:Fluorescent Dyes; D011090:Polyenes