segregation of fluorescent membrane lipids into distinct micrometric domains evidence for phase compartmentation of natural lipids荧光膜脂质隔离成不同的测微的域的证据相分隔的天然脂质.pdfVIP

Segregation of Fluorescent Membrane Lipids into Distinct Micrometric Domains: Evidence for Phase Compartmentation of Natural Lipids? 1 1 ´ ´ 2 1. Ludovic D9auria , Patrick Van Der Smissen , Frederic Bruyneel , Pierre J. Courtoy , Donatienne 1 . Tyteca * ´ ´ 1 CELL Unit, de Duve Institute and Universite catholique de Louvain, Brussels, Belgium, 2 CHOM Unit, Universite catholique de Louvain, Louvain-la-Neuve, Belgium Abstract Background: We recently reported that sphingomyelin (SM) analogs substituted on the alkyl chain by various fluorophores (e.g. BODIPY) readily inserted at trace levels into the plasma membrane of living erythrocytes or CHO cells and spontaneously concentrated into micrometric domains. Despite sharing the same fluorescent ceramide backbone, BODIPY- SM domains segregated from similar domains labelled by BODIPY-D-e-lactosylceramide (D-e-LacCer) and depended on endogenous SM. Methodology/Principal Findings: We show here that BODIPY-SM further differed from BODIPY-D-e-LacCer or - glucosylceramide (GlcCer) domains in temperature dependence, propensity to excimer formation, association with a glycosylphosphatidylinositol (GPI)-anchored fluorescent protein reporter, and lateral diffusion by FRAP, thus demonstrating different lipid phases and boundaries. Whereas BODIPY-D-e-LacCer behaved like BODIPY-GlcCer, its artificial stereoisomer, BODIPY-L-t-LacCer, behaved like BODIPY- and NBD-phosphatidylcholine (PC). Surprisingly, these t