Quaternary organization of the goodpasture autoantigen, the alpha 3(IV) collagen chain. Sequestration of two cryptic autoepitopes by intrapromoter interactions with the alpha4 and alpha5 NC1 domains.
Goodpasture's (GP) disease is caused by autoantibodies that target the alpha3(IV) collagen chain in the glomerular basement membrane (GBM). Goodpasture autoantibodies bind two conformational epitopes (E(A) and E(B)) located within the non-collagenous (NC1) domain of this chain, which are sequestered within the NC1 hexamer of the type IV collagen network containing the alpha3(IV), alpha4(IV), and alpha5(IV) chains. In this study, the quaternary organization of these chains and the molecular basis for the sequestration of the epitopes were investigated. This was accomplished by physicochemical and immunochemical characterization of the NC1 hexamers using chain-specific antibodies. The hexamers were found to have a molecular composition of (alpha3)(2)(alpha4)(2)(alpha5)(2) and to contain cross-linked alpha3-alpha5 heterodimers and alpha4-alpha4 homodimers. Together with association studies of individual NC1 domains, these findings indicate that the alpha3, alpha4, and alpha5 chains occur together in the same triple-helical protomer. In the GBM, this protomer dimerizes through NC1-NC1 domain interactions such that the alpha3, alpha4, and alpha5 chains of one protomer connect with the alpha5, alpha4, and alpha3 chains of the opposite protomer, respectively. The immunodominant Goodpasture autoepitope, located within the E(A) region, is sequestered within the alpha3alpha4alpha5 protomer near the triple-helical junction, at the interface between the alpha3NC1 and alpha5NC1 domains, whereas the E(B) epitope is sequestered at the interface between the alpha3NC1 and alpha4NC1 domains. The results also reveal the network distribution of the six chains of collagen IV in the renal glomerulus and provide a molecular explanation for the absence of the alpha3, alpha4, alpha5, and alpha6 chains in Alport syndrome.
Borza
Dorin-Bogdan
DB
Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee 37232, USA. Dorin-Bogdan.Borza@vanderbilt.edu
Bondar
Olga
O
Todd
Parvin
P
Sundaramoorthy
Munirathinam
M
Sado
Yoshikazu
Y
Ninomiya
Yoshifumi
Y
Hudson
Billy G
BG
D000911:Antibodies, Monoclonal / Q000378:metabolism; D001324:Autoantigens / Q000737:chemistry*; D001485:Basement Membrane / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D024141:Collagen Type IV / Q000737:chemistry*; D018360:Crystallography, X-Ray; D019281:Dimerization; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000378:metabolism; D008958:Models, Molecular; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011487:Protein Conformation; D020033:Protein Isoforms; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary
D000911:Antibodies, Monoclonal; D001324:Autoantigens; D024141:Collagen Type IV; D000939:Epitopes; D020033:Protein Isoforms; C057112:type IV collagen alpha3 chain