Transient surface delivery of invariant chain-MHC II complexes via endosomes: a quantitative study.
Newly synthesized major histocompatibility complex class II needs to be directed to late endocytic compartments to combine with peptide antigens. Efficient transport requires complexes of major histocompatibility complex class II and invariant chain (alphabetaIi). Since such complexes have been detected on the plasma membrane in human cells, this compartment was proposed as the primary destination for alphabetaIi exiting the trans-Golgi network. Here, I have used density gradient electrophoresis and selective biotinylation to investigate the trafficking route of alphabetaIi quantitatively. Density gradient electrophoresis analysis showed that alphabetaIi was transported from the trans-Golgi network to endosomes at approximately 1.7% min-1. Surface delivery of alphabetaIi was delayed relative to endosome transport by approximately 10 min and showed slower kinetics ( approximately 0.4% min-1), suggesting that alphabetaIi reached the plasma membrane only after arrival in endosomes. A biotinylation assay revealed that 20-40% of endosomal alphabetaIi was delivered to the plasma membrane at steady state, suggesting that surface alphabetaIi was entirely derived from endosomes. Surface alphabetaIi was rapidly re-internalized and either returned to the cell surface or accessed degradative compartments. Peptide loading commenced approximately 30 min after delivery to endosomes. Thus alphabetaIi directly traffics from trans-Golgi network to endosomes and enters an endosome-plasma membrane 'carousel' until transport to peptide-loading compartments ensues.
Lindner
Robert
R
Department of Cell Biology, Center of Anatomy, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. rli@zellbiologie.mh-hannover.de
D000818:Animals; D000938:Antigen-Presenting Cells / Q000378:metabolism; D001692:Biological Transport; D001710:Biotin / Q000378:metabolism; D019921:Biotinylation; D002462:Cell Membrane / Q000378:metabolism; D002469:Cell Separation; D004586:Electrophoresis; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism*; D005434:Flow Cytometry; D005802:Genes, MHC Class II; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D051379:Mice; D008808:Mice, Inbred CBA; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D010455:Peptides / Q000737:chemistry; D011233:Precipitin Tests; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors; D014407:Tumor Cells, Cultured
D010455:Peptides; D001710:Biotin