title: The goodpasture autoantigen. Identification of multiple cryptic epitopes on the NC1 domain of the alpha3(IV) collagen chain.
abstract: Goodpasture (GP) disease is an autoimmune disorder in which autoantibodies against the alpha3(IV) chain of type IV collagen bind to the glomerular and alveolar basement membranes, causing progressive glomerulonephritis and pulmonary hemorrhage. Two major conformational epitope regions have been identified on the noncollagenous domain of type IV collagen (NC1 domain) of the alpha3(IV) chain as residues 17-31 (E(A)) and 127-141 (E(B)) (Netzer, K.-O. et al. (1999) J. Biol. Chem. 274, 11267-11274). To determine whether these regions are two distinct epitopes or form a single epitope, three GP sera were fractionated by affinity chromatography on immobilized NC1 chimeras containing the E(A) and/or the E(B) region. Four subpopulations of GP antibodies with distinct epitope specificity for the alpha3(IV)NC1 domain were thus separated and characterized. They were designated GP(A), GP(B), GP(AB), and GP(X), to reflect their reactivity with E(A) only, E(B) only, both regions, and neither, respectively. Hence, regions E(A) and E(B) encompass critical amino acids that constitute three distinct epitopes for GP(A), GP(B), and GP(AB) antibodies, respectively, whereas the epitope for GP(X) antibodies is located in a different unknown region. The GP(A) antibodies were consistently immunodominant, accounting for 60-65% of the total immunoreactivity to alpha3(IV)NC1; thus, they probably play a major role in pathogenesis. Regions E(A) and E(B) are held in close proximity because they jointly form the epitope for Mab3, a monoclonal antibody that competes for binding with GP autoantibodies. All GP epitopes are sequestered in the hexamer configuration of the NC1 domain found in tissues and are inaccessible for antibody binding unless dissociation of the hexamer occurs, suggesting a possible mechanism for etiology of GP disease. GP antibodies have the capacity to extract alpha3(IV)NC1 monomers, but not dimers, from native human glomerular basement membrane hexamers, a property that may be of fundamental importance for the pathogenesis of the disease.
authors.affiliation: Department of Biochemistry, University of Kansas Medical Center, Kansas City, Kansas 66160, USA.
authors.forename: D B
authors.initials: DB
authors.lastname: Borza
authors.forename: K O
authors.initials: KO
authors.lastname: Netzer
authors.forename: A
authors.initials: A
authors.lastname: Leinonen
authors.forename: P
authors.initials: P
authors.lastname: Todd
authors.forename: J
authors.initials: J
authors.lastname: Cervera
authors.forename: J
authors.initials: J
authors.lastname: Saus
authors.forename: B G
authors.initials: BG
authors.lastname: Hudson
mesh_terms: D019867:Anti-Glomerular Basement Membrane Disease / Q000209:etiology / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000915:Antibody Affinity; D000918:Antibody Specificity; D001324:Autoantigens / Q000737:chemistry* / Q000302:isolation & purification; D001667:Binding, Competitive; D002460:Cell Line; D002846:Chromatography, Affinity; D003094:Collagen / Q000737:chemistry* / Q000302:isolation & purification; D024141:Collagen Type IV; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D008954:Models, Biological; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D001324:Autoantigens; D024141:Collagen Type IV; D000939:Epitopes; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; C057112:type IV collagen alpha3 chain; D003094:Collagen