title: Molecular cloning of the human Goodpasture antigen demonstrates it to be the alpha 3 chain of type IV collagen.
abstract: To characterize the autoantigen of Goodpasture's (anti-glomerular basement membrane) disease, a molecule of 26-kD reactive with autoantibodies from patients' sera was purified from collagenase digests of sheep glomerular basement membrane. Short internal amino acid sequences were obtained after tryptic or cyanogen bromide cleavage, and used to deduce redundant oligonucleotides for use in the polymerase chain reaction on cDNA derived from sheep renal cortex. Molecules of 175 bp were amplified and found to come from two cDNA sequences. One was identical to that of a type IV collagen chain (alpha 5) cloned from human placenta and shown to be expressed in human kidney. The other was from a type IV collagen chain with close similarities to alpha 1 and alpha 5 chains, and was used to obtain human cDNA sequences by cDNA library screening and by further polymerase chain reaction amplifications. The correspondence of the derived amino acid sequence of the new chain with published protein and cDNA sequences shows it to be the alpha 3 chain of type IV collagen. Its gene, COL4A3, maps to 2q36-2q37. The primary sequence and other characteristics of this chain confirm that it carries the Goodpasture antigen.
authors.affiliation: Department of Medicine, Royal Postgraduate Medical School, London, United Kingdom.
authors.forename: N
authors.initials: N
authors.lastname: Turner
authors.forename: P J
authors.initials: PJ
authors.lastname: Mason
authors.forename: R
authors.initials: R
authors.lastname: Brown
authors.forename: M
authors.initials: M
authors.lastname: Fox
authors.forename: S
authors.initials: S
authors.lastname: Povey
authors.forename: A
authors.initials: A
authors.lastname: Rees
authors.forename: C D
authors.initials: CD
authors.lastname: Pusey
mesh_terms: D000595:Amino Acid Sequence; D019867:Anti-Glomerular Basement Membrane Disease / Q000276:immunology*; D001324:Autoantigens / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D001485:Basement Membrane / Q000276:immunology; D015152:Blotting, Northern; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003094:Collagen / Q000032:analysis* / Q000235:genetics* / Q000302:isolation & purification; D024141:Collagen Type IV; D004247:DNA / Q000737:chemistry; D006801:Humans; D007678:Kidney Glomerulus / Q000276:immunology; D008969:Molecular Sequence Data
chemical_list: D001324:Autoantigens; D024141:Collagen Type IV; C057112:type IV collagen alpha3 chain; D003094:Collagen; D004247:DNA