A possible ligand of serum origin for the kidney autoantigen of Heymann nephritis.
Earlier studies have localized the Heymann nephritis (HN) autoantigen (gp330) in the coated pits of the plasma membrane and multivesicular bodies of the glomerular epithelial cell. Because of these locations in the glomerular epithelial cells, it has been suggested that the HN Ag may be a receptor. The aim of our study was to search for a ligand which can bind the HN autoantigen. Normal rat serum was subjected to SDS-PAGE under reducing and non-reducing conditions followed by Western analysis of the separated polypeptides. A reaction was revealed directly by autoradiography using 125I labeled HN autoantigen as a probe and indirectly by enzyme immunodetection using unlabeled nephritogenic autoantibody (anti-gp330) eluted from glomeruli of diseased rats followed by biotinylated rabbit anti-rat IgG avidin-peroxidase complex. A polypeptide of 76 kDa Mr was identified under non-reducing conditions as a serum protein reacting with the HN autoantigen. Reactivity of the 76-kDa polypeptide was lost when serum was electrophoresed under reducing conditions. Direct binding of the 76-kDa polypeptide obtained from serum to the HN autoantigen obtained from kidney suggests that the 76-kDa polypeptide may be a ligand for the autoantigen. This is the first documentation of a possible ligand for the HN autoantigen. Not only does this polypeptide bind to the HN autoantigen but it also shows direct binding with the nephritogenic autoantibody eluted from glomerular deposits. This characteristic of the 76-kDa polypeptide indicates that this serum protein may potentially play a role in the development of the glomerular lesion of active HN. Further analysis of this serum component should assist in understanding the normal function of the HN autoantigen.
Kanalas
J J
JJ
Department of Pediatrics, University of Texas Health Science Center, San Antonio 78284-7813.
Makker
S P
SP
D000818:Animals; D001323:Autoantibodies / Q000032:analysis; D001324:Autoantigens / Q000032:analysis* / Q000276:immunology; D001666:Binding Sites, Antibody; D001798:Blood Proteins / Q000302:isolation & purification / Q000378:metabolism*; D005921:Glomerulonephritis / Q000097:blood* / Q000276:immunology; D026541:Heymann Nephritis Antigenic Complex; D007678:Kidney Glomerulus / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000276:immunology / Q000378:metabolism*; D008970:Molecular Weight; D051381:Rats; D011917:Rats, Inbred Lew; D011971:Receptors, Immunologic / Q000302:isolation & purification / Q000378:metabolism*
D001323:Autoantibodies; D001324:Autoantigens; D001798:Blood Proteins; D026541:Heymann Nephritis Antigenic Complex; D008562:Membrane Glycoproteins; D011971:Receptors, Immunologic