Abstract
The antigen involved in the glomerulonephritis associated with antibodies to glomerular basement membrane (GBM) was purified from human GBM digested with highly purified clostridial collagenase. The purified nonreduced sample contained two components with closely similar mobilities on sodium dodecyl sulfate/polyacrylamide gel electrophoresis. After reduction they moved as one, nonantigenic, component, corresponding to a molecular weight of 26,000. Immunologically identical aggregates of higher molecular weight (i.e., 48,000) were also identified in the crude digest. Reduction of such aggregates after purification released some protein with a molecular weight of 26,000, but a large proportion was insensitive to reduction. Seven patients with Goodpasture syndrome all had circulating anti-GBM antibodies directed only against the purified antigen.
MeSH Terms
Amino Acids/analysis
Anti-Glomerular Basement Membrane Disease/immunology
Antigens, Surface/isolation & purification
Basement Membrane/immunology
Chromatography, Affinity
Enzyme-Linked Immunosorbent Assay
Humans
Kidney Glomerulus/immunology
Male
Microbial Collagenase
Molecular Weight
Chemicals
Amino Acids
Antigens, Surface
Microbial Collagenase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Wieslander J
Bygren P
Heinegård D
References (21)
21 references, click to expand
-
7-S collagen: characterization of an unusual basement membrane structure.
Eur J Biochem. 1980;108(1):239-50
PMID: 6250829
-
Entactin, a novel basal lamina-associated sulfated glycoprotein.
J Biol Chem. 1981 May 25;256(10):5209-14
PMID: 6262321
-
"Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.
Anal Biochem. 1981 Apr;112(2):195-203
PMID: 6266278
-
Partial characterization of newly synthesized proteoglycans isolated from the glomerular basement membrane.
J Cell Biol. 1981 Aug;90(2):527-32
PMID: 6793599
-
A network model for the organization of type IV collagen molecules in basement membranes.
Eur J Biochem. 1981 Nov;120(2):203-11
PMID: 6274634
-
Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity.
Anal Biochem. 1981 Nov 1;117(2):307-10
PMID: 6172996
-
Characterisation of human glomerular basement membrane antigenic fractions isolated by affinity chromatography utilising anti-glomerular basement membrane autoantibodies.
Biochem Biophys Res Commun. 1982 Feb 11;104(3):1025-32
PMID: 6280703
-
Antiglomerular basement membrane antibody: antibody specificity in different forms of glomerulonephritis.
Kidney Int. 1983 Jun;23(6):855-61
PMID: 6136625
-
STARCH-GEL ELECTROPHORESIS--APPLICATION TO THE CLASSIFICATION OF PITUITARY PROTEINS AND POLYPEPTIDES.
Metabolism. 1964 Oct;13:SUPPL:985-1002
PMID: 14228777
-
Studies on the renal glomerular basement membrane. Preparation and chemical composition.
J Biol Chem. 1967 Apr 25;242(8):1915-22
PMID: 6024780
-
Studies on the renal glomerular basement membrane. Nature of the carbohydrate units and their attachment to the peptide portion.
J Biol Chem. 1967 Apr 25;242(8):1923-32
PMID: 4290446
-
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature. 1970 Aug 15;227(5259):680-5
PMID: 5432063
-
Human glomerular basement membrane. Preparation and composition.
Biochemistry. 1970 Sep 15;9(19):3837-46
PMID: 4101157
-
Antigen identification in Goodpasture's syndrome.
Arch Pathol. 1971 Nov;92(5):329-33
PMID: 4938887
-
Isolation and immunological characterization of human glomerular basement membrane antigens.
Kidney Int. 1973 Feb;3(2):57-65
PMID: 4348613
-
Detection of anti-glomerular basement membrane antibodies by radioimmunological technique. Clinical application in human nephropathies.
J Clin Invest. 1974 Jul;54(1):128-37
PMID: 4857905
-
A simplified method for cyanogen bromide activation of agarose for affinity chromatography.
Anal Biochem. 1974 Jul;60(1):149-52
PMID: 4850305
-
The preparation of a bacterial collagenase containing negligible non-specific protease activity.
Prep Biochem. 1975;5(3):229-45
PMID: 170602
-
Laminin--a glycoprotein from basement membranes.
J Biol Chem. 1979 Oct 10;254(19):9933-7
PMID: 114518
-
Biochemistry and metabolism of basement membranes.
Int Rev Cytol. 1979;61:167-228
PMID: 389868
-
Study on the nature of the Goodpasture antigen using a basement membrane-producing mouse tumour.
Clin Exp Immunol. 1980 Mar;39(3):733-8
PMID: 6247110