title: Organization and dynamics of Fas transmembrane domain in raft membranes and modulation by ceramide.
abstract: To comprehend the molecular processes that lead to the Fas death receptor clustering in lipid rafts, a 21-mer peptide corresponding to its single transmembrane domain (TMD) was reconstituted into mammalian raft model membranes composed of an unsaturated glycerophospholipid, sphingomyelin, and cholesterol. The peptide membrane lateral organization and dynamics, and its influence on membrane properties, were studied by steady-state and time-resolved fluorescence techniques and by attenuated total reflection Fourier transformed infrared spectroscopy. Our results show that Fas TMD is preferentially localized in liquid-disordered membrane regions and undergoes a strong reorganization as the membrane composition is changed toward the liquid-ordered phase. This results from the strong hydrophobic mismatch between the length of the peptide hydrophobic stretch and the hydrophobic thickness of liquid-ordered membranes. The stability of nonclustered Fas TMD in liquid-disordered domains suggests that its sequence may have a protective function against nonligand-induced Fas clustering in lipid rafts. It has been reported that ceramide induces Fas oligomerization in lipid rafts. Here, it is shown that neither Fas TMD membrane organization nor its conformation is affected by ceramide. These results are discussed within the framework of Fas membrane signaling events.
authors.affiliation: Centro de Química Física-Molecular and Institute of Nanoscience and Nanotechnology, Instituto Superior Técnico, Lisbon, Portugal. bruno.castro@ist.utl.pt
authors.forename: Bruno M
authors.initials: BM
authors.lastname: Castro
authors.forename: Rodrigo F M
authors.initials: RF
authors.lastname: de Almeida
authors.forename: Erik
authors.initials: E
authors.lastname: Goormaghtigh
authors.forename: Aleksander
authors.initials: A
authors.lastname: Fedorov
authors.forename: Manuel
authors.initials: M
authors.lastname: Prieto
mesh_terms: D000595:Amino Acid Sequence; D017209:Apoptosis / Q000187:drug effects; D002518:Ceramides / Q000378:metabolism / Q000494:pharmacology*; D005454:Fluorescence Polarization; D021962:Membrane Microdomains / Q000187:drug effects* / Q000378:metabolism*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary / Q000187:drug effects; D021381:Protein Transport; D017550:Spectroscopy, Fourier Transform Infrared; D019014:fas Receptor / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D002518:Ceramides; C491901:FAS protein, human; D019014:fas Receptor