title: Using NK Cell Lipid Raft Fractionation to Understand the Role of Lipid Rafts in NK Cell Receptor Signaling.
abstract: Lipid rafts were first defined as detergent-resistant membranes (DRMs) due to their relative insolubility in non-ionic detergents. Although they should not be confused with lipid rafts, DRMs are a valuable starting point for the study of these membrane domains and the interactions of proteins with rafts.Here we describe the isolation of DRMs by ultracentrifugation on a sucrose gradient, a method we have used to study the role of lipid rafts in NKG2D-mediated signaling. We also describe raft fractionation of NK cells involving the selective solubility of β-octylglucoside (β-OG). OG is a non-ionic detergent that efficiently dissolves DRMs but does not disrupt protein associations with the cytoskeleton. Using these two techniques may yield useful information about the proteins involved in receptor recruitment into lipid rafts and the interactions of the actin cytoskeleton with lipid rafts.
authors.affiliation: Neurology Department, Hospital Universitario Central de Asturias, Oviedo, Asturias, Spain.
authors.forename: Esther
authors.initials: E
authors.lastname: Serrano-Pertierra
authors.affiliation: Immunology Department, Hospital Universitario Central de Asturias, Oviedo, Asturias, Spain. inmuno@hca.es.|Fundación Renal "Íñigo Álvarez de Toledo", Madrid, Spain. inmuno@hca.es.|Servicio de Inmunología, Avenida de Roma s/n, 33011, Oviedo, Asturias, Spain. inmuno@hca.es.
authors.forename: Carlos
authors.initials: C
authors.lastname: López-Larrea
mesh_terms: D002458:Cell Fractionation / Q000379:methods*; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D003902:Detergents / Q000737:chemistry; D005960:Glucosides / Q000737:chemistry; D006801:Humans; D007694:Killer Cells, Natural / Q000166:cytology / Q000378:metabolism*; D021962:Membrane Microdomains / Q000378:metabolism*; D055655:NK Cell Lectin-Like Receptor Subfamily K / Q000378:metabolism; D015398:Signal Transduction; D012995:Solubility
chemical_list: D003902:Detergents; D005960:Glucosides; C527085:KLRK1 protein, human; D055655:NK Cell Lectin-Like Receptor Subfamily K; C018619:octyl-beta-D-glucoside
keywords: DRM; Lipid raft; Membrane raft; NK cell; Sucrose gradient; Ultracentrifugation; β-octylglucoside