The assembly and stability of MHC class II-(alpha beta)2 superdimers.
X-ray crystallography of several MHC class II molecules revealed a structure described as a dimer of heterodimers, or a superdimer. This discovery led to the hypothesis that MHC class II molecules may interact with the TCR and CD4 as an (alpha beta)2 superdimer, potentially providing more stable and stimulatory interactions than can be provided by the simple alpha beta heterodimer alone. In this study, using chemical cross-linking, we provide evidence for the existence of the superdimers surface of B cells. We further characterize the superdimers and demonstrate that in lysates of B cells, I-Ek dimers and superdimers are derived from the same population of I-Ek molecules. Purified, I-Ek molecules in solution also exist as a mixture of 60-kDa dimers and 120-kDa superdimers, indicating that I-Ek has an intrinsic ability to form 120-kDa complexes in the absence of other cellular components. Peptide mapping showed that the alpha beta and (alpha beta)2 complexes are closely related and that the superdimers do not contain additional polypeptides not present in the dimers. The (alpha beta)2 complex displays thermal and pH stability similar to that of the alpha beta complex, both being denatured by SDS at temperatures above 50 degrees C and at a pH below 5. These data support the model that MHC class II has an intrinsic ability to assume the (alpha beta)2 superdimeric conformation, which may be important for interactions with the TCR and CD4 coreceptor.
Schafer
P H
PH
Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, IL 60208, USA.
Malapati
S
S
Hanfelt
K K
KK
Pierce
S K
SK
D000818:Animals; D001402:B-Lymphocytes / Q000737:chemistry* / Q000378:metabolism*; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D003432:Cross-Linking Reagents; D019281:Dimerization; D000949:Histocompatibility Antigens Class II / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D016393:Lymphoma, B-Cell; D046911:Macromolecular Substances; D051379:Mice; D008970:Molecular Weight; D011489:Protein Denaturation; D012967:Sodium Dodecyl Sulfate; D013696:Temperature; D014407:Tumor Cells, Cultured
D003432:Cross-Linking Reagents; D000949:Histocompatibility Antigens Class II; C024138:I-E-antigen; D046911:Macromolecular Substances; D012967:Sodium Dodecyl Sulfate