MHC class II molecules traffic into lipid rafts during intracellular transport.
There have been many studies demonstrating that a portion of MHC class II molecules reside in detergent-insoluble membrane domains (commonly referred to as lipid rafts). We have proposed that the function of raft association is to concentrate specific MHC class II-peptide complexes in plasma membrane microdomains that can facilitate efficient T cell activation. We now show that MHC class II becomes lipid raft associated before binding antigenic peptides. Using pulse-chase radiolabeling techniques, we find that newly synthesized MHC class II and MHC class II-invariant chain complexes initially reside in a detergent-soluble membrane fraction and acquire detergent insolubility as they traffic to lysosomal Ag processing compartments. Monensin, an inhibitor of protein transport through the Golgi apparatus, blocks association of newly synthesized MHC class II with lipid rafts. Treatment of cells with leupeptin, which inhibits invariant chain degradation, leads to the accumulation of MHC class II in lipid rafts within the lysosome-like Ag-processing compartments. Raft fractionation of lysosomal membranes confirmed the presence of MHC class II in detergent-insoluble microdomains in Ag-processing compartments. These findings indicate that newly synthesized MHC class II complexes are directed to detergent-insoluble lipid raft microdomains before peptide loading, a process that may facilitate the loading of similar peptides on MHC class II complexes in these microdomains.
Poloso
Neil J
NJ
Experimental Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Muntasell
Aura
A
Roche
Paul A
PA
D017951:Antigen Presentation / Q000276:immunology; D000944:Antigens, Differentiation, B-Lymphocyte / Q000737:chemistry / Q000378:metabolism; D002461:Cell Line, Transformed; D002592:Cetomacrogol; D003902:Detergents; D006681:HLA-D Antigens / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D000949:Histocompatibility Antigens Class II / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007424:Intracellular Fluid / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D008247:Lysosomes / Q000276:immunology / Q000378:metabolism; D046911:Macromolecular Substances; D021962:Membrane Microdomains / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000276:immunology; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D021381:Protein Transport / Q000276:immunology; D012995:Solubility; D013501:Surface-Active Agents
D000944:Antigens, Differentiation, B-Lymphocyte; D003902:Detergents; D006681:HLA-D Antigens; D000949:Histocompatibility Antigens Class II; D046911:Macromolecular Substances; D018832:Molecular Chaperones; D010446:Peptide Fragments; D021122:Protein Subunits; D013501:Surface-Active Agents; C045084:invariant chain; D002592:Cetomacrogol