1. Assembly of an abundant endogenous major histocompatibility complex class II/peptide complex in class II compartments
- Author
-
Stanislaw Morkowski, Hans J. Geuze, Monique J. Kleijmeer, Graça Raposo, and Alexander Y. Rudensky
- Subjects
Macromolecular Substances ,Immunology ,Peptide ,Biology ,Major histocompatibility complex ,Mice ,Antigens, CD ,Centrifugation, Density Gradient ,Immunology and Allergy ,Animals ,Humans ,Cells, Cultured ,Differential centrifugation ,chemistry.chemical_classification ,B-Lymphocytes ,HLA-D Antigens ,Membrane Glycoproteins ,Histocompatibility Antigens Class II ,Antibodies, Monoclonal ,Lysosome-Associated Membrane Glycoproteins ,Biological Transport ,Immunogold labelling ,Molecular biology ,Immunohistochemistry ,Precipitin Tests ,Recombinant Proteins ,beta-N-Acetylhexosaminidases ,Cell biology ,Cell Compartmentation ,Antigens, Differentiation, B-Lymphocyte ,Kinetics ,chemistry ,biology.protein ,Cell fractionation ,Peptides ,Percoll ,Alpha chain ,Intracellular ,Protein Binding - Abstract
To identify the intracellular site(s) of formation of an endogenous class II/peptide complex in a human B cell line, we employed kinetic pulse-chase labeling experiments followed by subcellular fractionation by Percoll density gradient centrifugation and immunogold labeling on ultrathin cryosections. For direct demonstration of assembly of such complexes, we used the monoclonal antibody YAe, which detects an endogenous complex of the mouse class II molecule I-Ab with a 17-amino acid peptide derived from the alpha chain of HLA-DR (DR alpha52-68). We show that in human B lymphocytes, these class II/peptide complexes assemble and transiently accumulate in major histocompatibility complex class II-enriched compartments before reaching the cell surface.
- Published
- 1997