Molecular cloning of a cDNA encoding a major pathogenic domain of the Heymann nephritis antigen gp330.
Heymann nephritis is an experimental autoimmune disease in rats that is characterized by accumulation of immune deposits (IDs) in kidney glomeruli. The disease is initiated by the binding of circulating antibodies to a membrane glycoprotein, gp330, which is a resident protein of clathrin-coated pits on glomerular epithelial cells (podocytes). We have defined a domain representing about 10% of gp330 that appears to be responsible for the formation of stable glomerular IDs. A cDNA clone (clone 14) was isolated from a rat kidney cDNA expression library by screening with IgG eluted from glomeruli of rats in early stages (3 days) of passive Heymann nephritis. The clone 14 cDNA contains an open reading frame encoding the C-terminal 319 amino acids of gp330. The predicted amino acid sequence contains four internal repeats of 11 amino acids, which are also found in the putative ligand-binding region of carbohydrate-binding lectin-like receptors. An antibody raised against the clone 14 fusion protein recognized gp330 by immunoblotting and induced formation of subepithelial IDs typical of passive Heymann nephritis when injected into normal rats. When the clone 14 fusion protein was used to immunize rats, subepithelial IDs of active Heymann nephritis were found after 12 weeks. No IDs were formed by active or passive immunization of rats with fusion proteins derived from other regions of gp330. These results demonstrate that clone 14 encodes a region of gp330 responsible for antibody binding and ID formation in vivo.
Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510.
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Pietromonaco
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Kerjaschki
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S
Binder
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R
Ullrich
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MG
Farquhar
D000595:Amino Acid Sequence; D000818:Animals; D001324:Autoantigens / Q000235:genetics*; D001327:Autoimmune Diseases / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D003488:Cyanogen Bromide; D004247:DNA / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D015723:Gene Library; D026541:Heymann Nephritis Antigenic Complex; D007074:Immunoglobulin G; D007678:Kidney Glomerulus / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000302:isolation & purification; D010449:Peptide Mapping; D051381:Rats; D012689:Sequence Homology, Nucleic Acid
D001324:Autoantigens; D026541:Heymann Nephritis Antigenic Complex; D007074:Immunoglobulin G; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D004247:DNA; D003488:Cyanogen Bromide