gp330 associates with a 44-kDa protein in the rat kidney to form the Heymann nephritis antigenic complex.
Using antibodies isolated from glomeruli of nephritic rats we have previously identified a 330-kDa cell surface glycoprotein (gp330) as a major pathogenic antigen of Heymann nephritis (HN), an experimental model of human membranous glomerulonephritis. Recently, we have isolated a cDNA clone, C14, encoding a polypeptide that contains a pathogenic epitope of HN responsible for the initiation of the disease. Subsequently, another protein, alpha 2-macroglobulin receptor-associated protein (alpha 2-MRAP), which is a subunit of the receptor for human alpha 2-macroglobulin/low density lipoprotein receptor-related protein (LRP), was shown to possess a high degree of sequence homology to the C14 protein (C14p). In this report, we have investigated the relationship between gp330, C14p, and alpha 2-MRAP. Immunoprecipitation studies demonstrate that gp330 forms a heterodimeric association with a 44-kDa polypeptide that is stable to detergent extraction and long-term centrifugation. Further, immunoblotting analysis on the purified complex indicates that the 44-kDa associated protein shares immunological identity to C14p and alpha 2-MRAP. In addition, antibodies eluted from glomeruli of HN rats and antibodies to a C14 fusion protein immunoprecipitated gp330 and the 44-kDa protein, demonstrating that the epitopes responsible for the initial events of HN are accessible within the complex. Based on these data, three models are proposed to explain how pathogenic epitopes in the gp330-44-kDa, HN antigenic complex may be presented at the cell surface and initiate the onset of HN.
Orlando
R A
RA
Division of Cellular and Molecular Medicine, University of California, San Diego, La Jolla 92093.
Kerjaschki
D
D
Kurihara
H
H
Biemesderfer
D
D
Farquhar
M G
MG
D000818:Animals; D000831:Animals, Newborn; D000906:Antibodies; D001324:Autoantigens / Q000378:metabolism; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D026541:Heymann Nephritis Antigenic Complex; D015151:Immunoblotting; D007668:Kidney / Q000276:immunology*; D007678:Kidney Glomerulus / Q000276:immunology* / Q000648:ultrastructure; D007687:Kidney Tubules, Proximal / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D008871:Microvilli / Q000276:immunology* / Q000648:ultrastructure; D008970:Molecular Weight; D011506:Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats
D000906:Antibodies; D001324:Autoantigens; D026541:Heymann Nephritis Antigenic Complex; D008562:Membrane Glycoproteins; D011506:Proteins