HLA-B27 in Transgenic Rats Forms Disulfide-Linked Heavy Chain Oligomers and Multimers That Bind to the Chaperone BiP
| Autor: | Tri M. Tran, Martha L. Dorris, Ekkehard May, Eiichi Furuta, Nimman Satumtira, Andrew Z. Wang, Joel D. Taurog |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
musculoskeletal diseases
Immunoprecipitation Transgene Immunology Mutant Endoplasmic Reticulum Animals Genetically Modified Pathogenesis HLA-B7 Antigen chemistry.chemical_compound Animals Immunology and Allergy Disulfides Transgenes skin and connective tissue diseases Endoplasmic Reticulum Chaperone BiP HLA-B27 Antigen Heat-Shock Proteins biology Molecular mass Chemistry Antibodies Monoclonal Molecular biology Rats Molecular Weight Protein Subunits Monomer Biochemistry Rats Inbred Lew Chaperone (protein) biology.protein Binding Sites Antibody Carrier Proteins Protein Processing Post-Translational Spleen Intracellular Molecular Chaperones Protein Binding |
| Zdroj: | The Journal of Immunology. 172:5110-5119 |
| ISSN: | 1550-6606 0022-1767 |
| DOI: | 10.4049/jimmunol.172.8.5110 |
| Popis: | To test the hypothesis that HLA-B27 predisposes to disease by forming disulfide-linked homodimers, we examined rats transgenic for HLA-B27, mutant Cys67Ser HLA-B27, or HLA-B7. In splenic Con A blasts from high transgene copy B27 lines that develop inflammatory disease, the anti-H chain mAb HC10 precipitated four bands of molecular mass 78–105 kDa and additional higher molecular mass material, seen by nonreducing SDS-PAGE. Upon reduction, all except one 78-kDa band resolved to 44 kDa, the size of the H chain monomer. The 78-kDa band was found to be BiP/Grp78, and the other high molecular mass material was identified as B27 H chain. Analysis of a disease-resistant low copy B27 line showed qualitatively similar high molecular mass bands that were less abundant relative to H chain monomer. Disease-prone rats with a Cys67Ser B27 mutant showed B27 H chain bands at 95 and 115 kDa and a BiP band at 78 kDa, whereas only scant high molecular mass bands were found in cells from control HLA-B7 rats. 125I-surface labeled B27 oligomers were immunoprecipitated with HC10, but not with a mAb to folded B27-β2-microglobulin-peptide complexes. Immunoprecipitation of BiP with anti-BiP Abs coprecipitated B27 H chain multimers. Folding and maturation of B27 were slow compared with B7. These data indicate that disulfide-linked intracellular H chain complexes are more prone to form and bind BiP in disease-prone wild-type B27 and B27-C67S rats than in disease-resistant HLA-B7 rats. The data support the hypothesis that accumulation of misfolded B27 participates in the pathogenesis of B27-associated disease. |
| Databáze: | OpenAIRE |
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