Dynamic association of CD45 with detergent-insoluble microdomains in T lymphocytes.
The receptor-like protein tyrosine phosphatase CD45 is essential for TCR signal transduction. Substrates of CD45 include the protein tyrosine kinases p56(lck) and p59(fyn), both of which have been shown to be enriched in detergent-insoluble microdomains. Here we find that there is a cholesterol-dependent association between CD45 and the raft-associated protein linker for activation of T cells, suggesting that CD45 and linker for activation of T cells may colocalize in lipid rafts. Consistent with this observation, we find that approximately 5% of total CD45 can be detected in Triton X-100-insoluble buoyant fractions of sucrose gradients, demonstrating that CD45 is not excluded from lipid rafts. Upon stimulation of T cells with anti-CD3, there is a reduction in the amount of CD45 found associating with lipid rafts. Our data suggest that CD45 is present in lipid rafts in T cells before activation, perhaps to activate raft-associated p56(lck), allowing membrane-proximal signaling events to proceed. Furthermore, the reduction in CD45 content of lipid rafts after CD3 stimulation may serve to limit the amounts of activated p56(lck) in rafts and thus possibly the duration of T cell responses.
Edmonds
Stuart D
SD
Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Canada.
Ostergaard
Hanne L
HL
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D002465:Cell Movement / Q000276:immunology; D002592:Cetomacrogol; D002999:Clone Cells; D003505:Cyclodextrins / Q000494:pharmacology; D003593:Cytoplasm / Q000276:immunology / Q000378:metabolism; D003902:Detergents; D015536:Down-Regulation / Q000187:drug effects; D017493:Leukocyte Common Antigens / Q000378:metabolism* / Q000502:physiology; D008213:Lymphocyte Activation; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000378:metabolism; D021962:Membrane Microdomains / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism* / Q000502:physiology; D008565:Membrane Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D017830:Octoxynol; D010750:Phosphoproteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D010766:Phosphorylation; D010938:Plant Oils; D011092:Polyethylene Glycols; D012995:Solubility; D013601:T-Lymphocytes / Q000187:drug effects / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism; D047392:beta-Cyclodextrins
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D003505:Cyclodextrins; D003902:Detergents; C491734:Lat protein, mouse; D008565:Membrane Proteins; D010750:Phosphoproteins; D010938:Plant Oils; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D011092:Polyethylene Glycols; D014443:Tyrosine; D017830:Octoxynol; D002592:Cetomacrogol; C033084:polyethylene glycol oleyl ether; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D017493:Leukocyte Common Antigens