title: Folding and assembly of a human MHC class II molecule in a cell-free system.
abstract: The assembly of a human major histocompatibility complex (MHC) class II molecule was investigated in a cell-free system capable of the synthesis, sequestration, and processing of the protein chains. As assessed by the conformation-sensitive monoclonal antibody L243, the formation of HLA-DR alpha/beta heterodimer required cotranslation of alpha and beta mRNA in the presence of both oxidized glutathione and canine pancreas endoplasmic reticulum (ER) vesicles. The assembly of alpha/beta dimer could also be initiated by the post-translational addition of oxidized glutathione. Using the post-translational assay system, we investigated the effect of the depletion of ER lumenal content proteins on the folding and assembly of the MHC class II chains. Both the rate and extent of folding of alpha chain and beta chain and the post-translational assembly of alpha/beta dimer is greatly reduced in the depleted ER vesicles. Conversely, the extent of aggregate formation is increased. Upon reconstitution of the depleted ER vesicles with lumenal proteins, the folding of alpha chain is accelerated and the assembly of alpha/beta dimer is increased.
authors.affiliation: Department of Molecular Microbiology and Immunology, St. Louis University School of Medicine, MO 63104, USA.
authors.forename: D
authors.initials: D
authors.lastname: Qu
authors.forename: M
authors.initials: M
authors.lastname: Green
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002474:Cell-Free System; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000737:chemistry; D005978:Glutathione / Q000031:analogs & derivatives / Q000737:chemistry; D019803:Glutathione Disulfide; D006684:HLA-DR Antigens / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D010084:Oxidation-Reduction; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D006684:HLA-DR Antigens; D011506:Proteins; D005978:Glutathione; D019803:Glutathione Disulfide