Analysis of lipid rafts in T cells.
The plasma membrane of T cells is made of a combination of glycosphingolipids and protein receptors organized in glycolipoprotein microdomains termed lipid rafts. The structural assembly of lipid rafts was investigated by various physical and biochemical assays. Depending on the differentiation status of T cells, the lipid rafts seclude various protein receptors involved in T cell signaling, cytoskeleton reorganization, membrane trafficking, and the entry of infectious organisms into the cells. This review article summarizes the most common methods, and their limits and advantages for analyzing the composition and assembly of lipid rafts with protein receptors into lipid rafts microdomains in plasma membrane of T cells. It also includes new methods such as ELISA/Polysorp and flow cytometry, and a combined sucrose gradient centrifugation-FPLC-Western blot strategy developed in our laboratory to study non-covalent interactions between the GM1 glycosphingolipid and protein receptors in plasma membrane of T cells.
Thomas
Sunil
S
Department of Microbiology, Mount Sinai School of Medicine, 1 Gustave L. Levy Place, New York, NY 10029, USA.
Preda-Pais
Anca
A
Casares
Sofia
S
Brumeanu
Teodor-D
TD
D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation; D002855:Chromatography, Thin Layer; D003599:Cytoskeleton / Q000648:ultrastructure; D005434:Flow Cytometry; D006028:Glycosphingolipids / Q000302:isolation & purification / Q000502:physiology; D006801:Humans; D008024:Ligands; D046911:Macromolecular Substances; D008563:Membrane Lipids / Q000302:isolation & purification / Q000502:physiology; D021962:Membrane Microdomains / Q000502:physiology*; D008565:Membrane Proteins / Q000302:isolation & purification / Q000502:physiology; D008856:Microscopy, Fluorescence; D011956:Receptors, Cell Surface / Q000502:physiology; D013601:T-Lymphocytes / Q000276:immunology / Q000648:ultrastructure*
D006028:Glycosphingolipids; D008024:Ligands; D046911:Macromolecular Substances; D008563:Membrane Lipids; D008565:Membrane Proteins; D011956:Receptors, Cell Surface