Distinct differences in association of MHC class I with endoplasmic reticulum proteins in wild-type, and beta 2-microglobulin- and TAP-deficient cell lines.
In this study we have compared the interaction of human MHC class I molecules with IgG heavy chain (HC) binding protein (BiP), calnexin, calreticulin, tapasin and TAP in beta(2)-microglobulin (beta(2)m)- or TAP-deficient cells, as well as in wild-type B-LCL cells. Distinct differences between the association of HC and these endoplasmic reticulum (ER) proteins were found in the three cell lines. In the absence of beta(2)m (Daudi cells), HC associated with both BiP and calnexin. A prominent portion of HC was complexed simultaneously to both chaperones, as indicated by co-precipitation with either anti-calnexin or anti-class I antisera. In the presence of beta(2)m, but absence of TAP (T2 cells), HC could be co-precipitated with calnexin, whereas no detectable interaction with BiP could be demonstrated. This suggests that calnexin interacts with HC at a later stage than BiP. In B-LCL cells, HC-beta(2)m associated with calreticulin and tapasin, whereas no interaction with calnexin and BiP was observed. In the absence of beta(2)m, HC were rapidly degraded in the ER, while the ER retained HC were stabilized in the presence of beta(2)m, even in the absence of TAP. The dissociation of class I molecules from TAP in B-LCL cells correlated with the kinetics of appearance of class I molecules on the cell surface, suggesting that TAP retains peptide-free class I molecules in the ER. Taken together, our results suggest the model that BiP and calnexin sequentially control the folding of MHC class I, before MHC class I molecules associate with the loading complex.
Paulsson
K M
KM
Tumor Immunology, Lund University, Solvegatan 21, 22362 Lund, Sweden.
Wang
P
P
Anderson
P O
PO
Chen
S
S
Pettersson
R F
RF
Li
S
S
D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D000071450:ATP Binding Cassette Transporter, Subfamily B, Member 3; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism* / Q000502:physiology; D017920:Antiporters / Q000378:metabolism; D001692:Biological Transport / Q000276:immunology; D002135:Calcium-Binding Proteins / Q000378:metabolism; D037281:Calnexin; D037282:Calreticulin; D002352:Carrier Proteins / Q000378:metabolism; D019281:Dimerization; D004721:Endoplasmic Reticulum / Q000276:immunology / Q000378:metabolism*; D000091342:Endoplasmic Reticulum Chaperone BiP; D006680:HLA Antigens / Q000378:metabolism*; D006360:Heat-Shock Proteins; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D007136:Immunoglobulins / Q000378:metabolism; D026901:Membrane Transport Proteins; D018832:Molecular Chaperones / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism; D014407:Tumor Cells, Cultured; D001613:beta 2-Microglobulin / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D000071450:ATP Binding Cassette Transporter, Subfamily B, Member 3; D018528:ATP-Binding Cassette Transporters; D017920:Antiporters; D002135:Calcium-Binding Proteins; D037282:Calreticulin; D002352:Carrier Proteins; D000091342:Endoplasmic Reticulum Chaperone BiP; D006680:HLA Antigens; D006360:Heat-Shock Proteins; D015395:Histocompatibility Antigens Class I; D007074:Immunoglobulin G; D007136:Immunoglobulins; D026901:Membrane Transport Proteins; D018832:Molecular Chaperones; D012261:Ribonucleoproteins; C073119:TAP1 protein, human; D001613:beta 2-Microglobulin; C107762:tapasin; D037281:Calnexin; C071692:TAP2 protein, human