RRC ID 36036
Author Kiyokawa E, Baba T, Otsuka N, Makino A, Ohno S, Kobayashi T.
Title Spatial and functional heterogeneity of sphingolipid-rich membrane domains.
Journal J. Biol. Chem.
Abstract Little is known about the organization of lipids in biomembranes. Lipid rafts are defined as sphingolipid- and cholesterol-rich clusters in the membrane. Details of the lipid distribution of lipid rafts are not well characterized mainly because of a lack of appropriate probes. Ganglioside GM1-specific protein, cholera toxin, has long been the only lipid probe of lipid rafts. Recently it was shown that earthworm toxin, lysenin, specifically recognizes sphingomyelin-rich membrane domains. Binding of lysenin to sphingomyelin is accompanied by the oligomerization of the toxin that leads to pore formation in the target membrane. In this study, we generated a truncated lysenin mutant that does not oligomerize and thus is non-toxic. Using this mutant lysenin, we showed that plasma membrane sphingomyelin-rich domains are spatially distinct from ganglioside GM1-rich membrane domains in Jurkat T cells. Like T cell receptor activation and cross-linking of GM1, cross-linking of sphingomyelin induced calcium influx and ERK phosphorylation in the cell. However, unlike CD3 or GM1, cross-linking of sphingomyelin did not induce significant protein tyrosine phosphorylation. Combination of lysenin and sphingomyelinase treatment suggested the involvement of G-protein-coupled receptor in sphingomyelin-mediated signal transduction. These results thus suggest that the sphingomyelin-rich domain provides a functional signal cascade platform that is distinct from those provided by T cell receptor or GM1. Our study therefore elucidates the spatial and functional heterogeneity of lipid rafts.
Volume 280(25)
Pages 24072-84
Published 2005-6-24
DOI 10.1074/jbc.M502244200
PII M502244200
PMID 15840575
MeSH Cell Line Flow Cytometry Fluorescent Antibody Technique G(M1) Ganglioside / metabolism* Humans Mutagenesis Proteins / genetics Proteins / metabolism Recombinant Proteins / genetics Recombinant Proteins / metabolism Sequence Deletion Sphingomyelin Phosphodiesterase / genetics Sphingomyelin Phosphodiesterase / metabolism Toxins, Biological
IF 4.106
Times Cited 111
DNA material pQE30/His6-Venus-NT-Lys (RDB13959) pQE30/His6-mRFP-NT-Lys (RDB13960)