title: Raft coalescence and FcγRIIA activation upon sphingomyelin clustering induced by lysenin.
abstract: Activation of immunoreceptor FcγRIIA by cross-linking with antibodies is accompanied by coalescence of sphingolipid/cholesterol-rich membrane rafts leading to the formation of signaling platforms of the receptor. In this report we examined whether clustering of the raft lipid sphingomyelin can reciprocally induce partition of FcγRIIA to rafts. To induce sphingomyelin clustering, cells were exposed to non-lytic concentrations of GST-lysenin which specifically recognizes sphingomyelin. The lysenin/sphingomyelin complexes formed microscale assemblies composed of GST-lysenin oligomers engaging sphingomyelin of rafts. Upon sphingomyelin clustering, non-cross-linked FcγRIIA associated with raft-derived detergent-resistant membrane fractions as revealed by density gradient centrifugation. Pretreatment of cells with GST-lysenin also increased the size of detergent-insoluble molecular complexes of activated FcγRIIA. Sphingomyelin clustering triggered tyrosine phosphorylation of the receptor and its accompanying proteins, Cbl and NTAL, in the absence of receptor ligands and enhanced phosphorylation of these proteins in the ligand presence. These data indicate that clustering of plasma membrane sphingomyelin induces coalescence of rafts and triggers signaling events analogous to those caused by FcγRIIA activation.
authors.affiliation: Department of Cell Biology, Nencki Institute of Experimental Biology, Warsaw, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Kulma
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kwiatkowska
authors.forename: Andrzej
authors.initials: A
authors.lastname: Sobota
mesh_terms: D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D005982:Glutathione Transferase / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D010766:Phosphorylation; D017452:Receptors, IgG / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013109:Sphingomyelins / Q000378:metabolism*; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism; D014118:Toxins, Biological / Q000737:chemistry / Q000378:metabolism*; D020298:U937 Cells
chemical_list: C080568:Fc gamma receptor IIA; D017452:Receptors, IgG; D011994:Recombinant Proteins; D013109:Sphingomyelins; D014118:Toxins, Biological; C107040:lysenin; D005982:Glutathione Transferase