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The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1992 Feb;89(2):592–601. doi: 10.1172/JCI115625

Molecular cloning of the human Goodpasture antigen demonstrates it to be the alpha 3 chain of type IV collagen.

N Turner 1, P J Mason 1, R Brown 1, M Fox 1, S Povey 1, A Rees 1, C D Pusey 1
PMCID: PMC442892  PMID: 1737849

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.

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Selected References

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  1. Barker D. F., Hostikka S. L., Zhou J., Chow L. T., Oliphant A. R., Gerken S. C., Gregory M. C., Skolnick M. H., Atkin C. L., Tryggvason K. Identification of mutations in the COL4A5 collagen gene in Alport syndrome. Science. 1990 Jun 8;248(4960):1224–1227. doi: 10.1126/science.2349482. [DOI] [PubMed] [Google Scholar]
  2. Birnstiel M. L., Busslinger M., Strub K. Transcription termination and 3' processing: the end is in site! Cell. 1985 Jun;41(2):349–359. doi: 10.1016/s0092-8674(85)80007-6. [DOI] [PubMed] [Google Scholar]
  3. Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
  4. Bowman C., Lockwood C. M. Clinical application of a radio-immunoassay for auto-antibodies to glomerular basement membrane. J Clin Lab Immunol. 1985 Aug;17(4):197–202. [PubMed] [Google Scholar]
  5. Butkowski R. J., Langeveld J. P., Wieslander J., Hamilton J., Hudson B. G. Localization of the Goodpasture epitope to a novel chain of basement membrane collagen. J Biol Chem. 1987 Jun 5;262(16):7874–7877. [PubMed] [Google Scholar]
  6. Butkowski R. J., Shen G. Q., Wieslander J., Michael A. F., Fish A. J. Characterization of type IV collagen NC1 monomers and Goodpasture antigen in human renal basement membranes. J Lab Clin Med. 1990 Mar;115(3):365–373. [PubMed] [Google Scholar]
  7. Butkowski R. J., Wieslander J., Kleppel M., Michael A. F., Fish A. J. Basement membrane collagen in the kidney: regional localization of novel chains related to collagen IV. Kidney Int. 1989 May;35(5):1195–1202. doi: 10.1038/ki.1989.110. [DOI] [PubMed] [Google Scholar]
  8. Bygren P., Wieslander J., Heinegärd D. Glomerulonephritis induced in sheep by immunization with human glomerular basement membrane. Kidney Int. 1987 Jan;31(1):25–31. doi: 10.1038/ki.1987.4. [DOI] [PubMed] [Google Scholar]
  9. Cashman S. J., Pusey C. D., Evans D. J. Extraglomerular distribution of immunoreactive Goodpasture antigen. J Pathol. 1988 May;155(1):61–70. doi: 10.1002/path.1711550110. [DOI] [PubMed] [Google Scholar]
  10. Derry C. J., Dunn M. J., Rees A. J., Pusey C. D. Restricted specificity of the autoantibody response in Goodpasture's syndrome demonstrated by two-dimensional western blotting. Clin Exp Immunol. 1991 Dec;86(3):457–463. doi: 10.1111/j.1365-2249.1991.tb02953.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Edwards Y. H., Parkar M., Povey S., West L. F., Parrington J. M., Solomon E. Human myosin heavy chain genes assigned to chromosome 17 using a human cDNA clone as probe. Ann Hum Genet. 1985 May;49(Pt 2):101–109. doi: 10.1111/j.1469-1809.1985.tb01681.x. [DOI] [PubMed] [Google Scholar]
  12. Gunwar S., Saus J., Noelken M. E., Hudson B. G. Glomerular basement membrane. Identification of a fourth chain, alpha 4, of type IV collagen. J Biol Chem. 1990 Apr 5;265(10):5466–5469. [PubMed] [Google Scholar]
  13. Hostikka S. L., Eddy R. L., Byers M. G., Höyhtyä M., Shows T. B., Tryggvason K. Identification of a distinct type IV collagen alpha chain with restricted kidney distribution and assignment of its gene to the locus of X chromosome-linked Alport syndrome. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1606–1610. doi: 10.1073/pnas.87.4.1606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hostikka S. L., Tryggvason K. The complete primary structure of the alpha 2 chain of human type IV collagen and comparison with the alpha 1(IV) chain. J Biol Chem. 1988 Dec 25;263(36):19488–19493. [PubMed] [Google Scholar]
  15. Hudson B. G., Wieslander J., Wisdom B. J., Jr, Noelken M. E. Goodpasture syndrome: molecular architecture and function of basement membrane antigen. Lab Invest. 1989 Sep;61(3):256–269. [PubMed] [Google Scholar]
  16. Jeraj K., Michael A. F., Fish A. J. Immunologic similarities between Goodpasture's and Steblay's antibodies. Clin Immunol Immunopathol. 1982 May;23(2):408–413. doi: 10.1016/0090-1229(82)90125-8. [DOI] [PubMed] [Google Scholar]
  17. Kleppel M. M., Michael A. F., Fish A. J. Antibody specificity of human glomerular basement membrane type IV collagen NC1 subunits. Species variation in subunit composition. J Biol Chem. 1986 Dec 15;261(35):16547–16552. [PubMed] [Google Scholar]
  18. Kleppel M. M., Michael A. F., Fish A. J. Antibody specificity of human glomerular basement membrane type IV collagen NC1 subunits. Species variation in subunit composition. J Biol Chem. 1986 Dec 15;261(35):16547–16552. [PubMed] [Google Scholar]
  19. Kleppel M. M., Santi P. A., Cameron J. D., Wieslander J., Michael A. F. Human tissue distribution of novel basement membrane collagen. Am J Pathol. 1989 Apr;134(4):813–825. [PMC free article] [PubMed] [Google Scholar]
  20. Langeveld J. P., Wieslander J., Timoneda J., McKinney P., Butkowski R. J., Wisdom B. J., Jr, Hudson B. G. Structural heterogeneity of the noncollagenous domain of basement membrane collagen. J Biol Chem. 1988 Jul 25;263(21):10481–10488. [PubMed] [Google Scholar]
  21. LeFranc M. P., Forster A., Baer R., Stinson M. A., Rabbitts T. H. Diversity and rearrangement of the human T cell rearranging gamma genes: nine germ-line variable genes belonging to two subgroups. Cell. 1986 Apr 25;45(2):237–246. doi: 10.1016/0092-8674(86)90388-0. [DOI] [PubMed] [Google Scholar]
  22. Lerner R. A., Glassock R. J., Dixon F. J. The role of anti-glomerular basement membrane antibody in the pathogenesis of human glomerulonephritis. J Exp Med. 1967 Dec 1;126(6):989–1004. doi: 10.1084/jem.126.6.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lichter P., Tang C. J., Call K., Hermanson G., Evans G. A., Housman D., Ward D. C. High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones. Science. 1990 Jan 5;247(4938):64–69. doi: 10.1126/science.2294592. [DOI] [PubMed] [Google Scholar]
  24. MacDonald R. J., Swift G. H., Przybyla A. E., Chirgwin J. M. Isolation of RNA using guanidinium salts. Methods Enzymol. 1987;152:219–227. doi: 10.1016/0076-6879(87)52023-7. [DOI] [PubMed] [Google Scholar]
  25. Marston F. A. The purification of eukaryotic polypeptides synthesized in Escherichia coli. Biochem J. 1986 Nov 15;240(1):1–12. doi: 10.1042/bj2400001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. McCoy R. C., Johnson H. K., Stone W. J., Wilson C. B. Absence of nephritogenic GBM antigen(s) in some patients with hereditary nephritis. Kidney Int. 1982 Apr;21(4):642–652. doi: 10.1038/ki.1982.72. [DOI] [PubMed] [Google Scholar]
  27. Morrison K. E., Germino G. G., Reeders S. T. Use of the polymerase chain reaction to clone and sequence a cDNA encoding the bovine alpha 3 chain of type IV collagen. J Biol Chem. 1991 Jan 5;266(1):34–39. [PubMed] [Google Scholar]
  28. Pihlajaniemi T., Tryggvason K., Myers J. C., Kurkinen M., Lebo R., Cheung M. C., Prockop D. J., Boyd C. D. cDNA clones coding for the pro-alpha1(IV) chain of human type IV procollagen reveal an unusual homology of amino acid sequences in two halves of the carboxyl-terminal domain. J Biol Chem. 1985 Jun 25;260(12):7681–7687. [PubMed] [Google Scholar]
  29. Purdue P. E., Lumb M. J., Fox M., Griffo G., Hamon-Benais C., Povey S., Danpure C. J. Characterization and chromosomal mapping of a genomic clone encoding human alanine:glyoxylate aminotransferase. Genomics. 1991 May;10(1):34–42. doi: 10.1016/0888-7543(91)90481-s. [DOI] [PubMed] [Google Scholar]
  30. Pusey C. D., Dash A., Kershaw M. J., Morgan A., Reilly A., Rees A. J., Lockwood C. M. A single autoantigen in Goodpasture's syndrome identified by a monoclonal antibody to human glomerular basement membrane. Lab Invest. 1987 Jan;56(1):23–31. [PubMed] [Google Scholar]
  31. Pöschl E., Pollner R., Kühn K. The genes for the alpha 1(IV) and alpha 2(IV) chains of human basement membrane collagen type IV are arranged head-to-head and separated by a bidirectional promoter of unique structure. EMBO J. 1988 Sep;7(9):2687–2695. doi: 10.1002/j.1460-2075.1988.tb03122.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rees A. J., Peters D. K., Amos N., Welsh K. I., Batchelor J. R. The influence of HLA-linked genes on the severity of anti-GBM antibody-mediated nephritis. Kidney Int. 1984 Oct;26(4):445–450. doi: 10.1038/ki.1984.194. [DOI] [PubMed] [Google Scholar]
  33. Sarkar G., Sommer S. S. Access to a messenger RNA sequence or its protein product is not limited by tissue or species specificity. Science. 1989 Apr 21;244(4902):331–334. doi: 10.1126/science.2565599. [DOI] [PubMed] [Google Scholar]
  34. Saus J., Wieslander J., Langeveld J. P., Quinones S., Hudson B. G. Identification of the Goodpasture antigen as the alpha 3(IV) chain of collagen IV. J Biol Chem. 1988 Sep 15;263(26):13374–13380. [PubMed] [Google Scholar]
  35. Savage C. O., Pusey C. D., Kershaw M. J., Cashman S. J., Harrison P., Hartley B., Turner D. R., Cameron J. S., Evans D. J., Lockwood C. M. The Goodpasture antigen in Alport's syndrome: studies with a monoclonal antibody. Kidney Int. 1986 Jul;30(1):107–112. doi: 10.1038/ki.1986.158. [DOI] [PubMed] [Google Scholar]
  36. Saxena R., Isaksson B., Bygren P., Wieslander J. A rapid assay for circulating anti-glomerular basement membrane antibodies in Goodpasture syndrome. J Immunol Methods. 1989 Mar 10;118(1):73–78. doi: 10.1016/0022-1759(89)90055-0. [DOI] [PubMed] [Google Scholar]
  37. Segelmark M., Butkowski R., Wieslander J. Antigen restriction and IgG subclasses among anti-GBM autoantibodies. Nephrol Dial Transplant. 1990;5(12):991–996. doi: 10.1093/ndt/5.12.991. [DOI] [PubMed] [Google Scholar]
  38. Siebold B., Deutzmann R., Kühn K. The arrangement of intra- and intermolecular disulfide bonds in the carboxyterminal, non-collagenous aggregation and cross-linking domain of basement-membrane type IV collagen. Eur J Biochem. 1988 Oct 1;176(3):617–624. doi: 10.1111/j.1432-1033.1988.tb14321.x. [DOI] [PubMed] [Google Scholar]
  39. Soininen R., Huotari M., Hostikka S. L., Prockop D. J., Tryggvason K. The structural genes for alpha 1 and alpha 2 chains of human type IV collagen are divergently encoded on opposite DNA strands and have an overlapping promoter region. J Biol Chem. 1988 Nov 25;263(33):17217–17220. [PubMed] [Google Scholar]
  40. Spiro R. G. Studies on the renal glomerular basement membrane. Preparation and chemical composition. J Biol Chem. 1967 Apr 25;242(8):1915–1922. [PubMed] [Google Scholar]
  41. Steblay R. W., Rudofsky U. H. Experimental autoimmune glomerulonephritis induced by anti-glomerular basement membrane antibody. II. Effects of injecting heterologous, homologous, or autologous glomerular basement membranes and complete Freund's adjuvant into sheep. Am J Pathol. 1983 Nov;113(2):125–133. [PMC free article] [PubMed] [Google Scholar]
  42. Wieslander J., Kataja M., Hudson B. G. Characterization of the human Goodpasture antigen. Clin Exp Immunol. 1987 Aug;69(2):332–340. [PMC free article] [PubMed] [Google Scholar]
  43. Wieslander J., Kataja M., Hudson B. G. Characterization of the human Goodpasture antigen. Clin Exp Immunol. 1987 Aug;69(2):332–340. [PMC free article] [PubMed] [Google Scholar]
  44. Wong Z., Wilson V., Patel I., Povey S., Jeffreys A. J. Characterization of a panel of highly variable minisatellites cloned from human DNA. Ann Hum Genet. 1987 Oct;51(Pt 4):269–288. doi: 10.1111/j.1469-1809.1987.tb01062.x. [DOI] [PubMed] [Google Scholar]

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