Beta 2-microglobulin and calnexin can independently promote folding and disulfide bond formation in class I histocompatibility proteins.
Class I histocompatibility proteins fold and assemble with beta 2-microglobulin (beta 2m) into heterodimers before binding short peptides in the endoplasmic reticulum. Here, we show that class I proteins rapidly form disulfide bonds, and that the process is highly reversible in Daudi cells lacking beta 2m. Three distinct class I protein conformations are present in equal amounts in these cells, each associated with the molecular chaperone calnexin. When binding of calnexin is inhibited by the glucosidase inhibitor castanospermine, fully oxidized class I proteins are no longer detected, suggesting that calnexin is required for completion of folding. However, in Daudi cells transfected to express beta 2m, castanospermine decreases only slightly the levels of fully oxidized class I proteins, indicating that folding is much less dependent on calnexin in the presence of beta 2m. Furthermore, calreticulin, a chaperone with functional similarities to calnexin, associates with class I molecules in beta 2m-positive cells. but not in Daudi cells, consistent with completion of folding and disulfide bond formation of class I heavy chains before binding to calreticulin occurs. This study demonstrates that calnexin and beta 2m can function independently to promote folding of class I heavy chains prior to formation of stable class I dimers.
Tector
M
M
Department of Pathology, University of Pittsburgh School of Medicine, PA 15213, U.S.A.
Zhang
Q
Q
Salter
R D
RD
D002051:Burkitt Lymphoma / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D037281:Calnexin; D037282:Calreticulin; D002453:Cell Cycle / Q000276:immunology; D004220:Disulfides / Q000378:metabolism*; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D018832:Molecular Chaperones / Q000502:physiology*; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D017510:Protein Folding; D012261:Ribonucleoproteins / Q000378:metabolism; D014407:Tumor Cells, Cultured; D001613:beta 2-Microglobulin / Q000502:physiology*
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D004220:Disulfides; D015395:Histocompatibility Antigens Class I; D018832:Molecular Chaperones; D010455:Peptides; D012261:Ribonucleoproteins; D001613:beta 2-Microglobulin; D037281:Calnexin