Identification of antigenic epitopes in type IV collagen by use of synthetic peptides.
Peptides representing potential antigenic regions of the NC-1 and 7-S domains of the human alpha 1 and alpha 2, and bovine alpha 3 chains of type IV collagen were synthesized either chemically or by the recombinant DNA technique and tested by ELISA using antibodies raised in rabbits against the whole type IV collagen or the NC-1 domain. Sera from patients with Goodpasture syndrome (GP) or with acute poststreptococcal glomerulonephritis (APSGN) were also tested. The location of antigenic determinants was predicted from the primary and secondary structure of the chains, that is, aromaticity, hydrophilicity and presence of beta-turns. All synthetic peptides reacted with the antiserum to type IV collagen (anti-Col IV). Whereas all peptides arising from the NC-1 domain reacted with anti-NC-1, intact 7-S or peptides of the alpha 1 or alpha 2 chain of the 7-S domain did not react. However intact 7-S reacted with anti-Col IV. Two synthetic peptides from the NC-1 domain of alpha 1, (a.a. 71-90 and a.a. 176-190), one from the alpha 2 (a.a. 70-83) and four from the alpha 3 chain (a.a. 72-89, a.a. 104-117, a.a. 133-145, a.a. 185-203) reacted with anti-NC-1 and anti-COL IV. The above peptides, except alpha 3 (72-89) and alpha 3 (185-203), were tested and found to be reactive with sera from patients with GP.(ABSTRACT TRUNCATED AT 250 WORDS)
Kefalides
N A
NA
Connective Tissue Research Institute, School of Dental Medicine, University of Pennsylvania, Philadelphia.
Ohno
N
N
Wilson
C B
CB
Fillit
H
H
Zabriski
J
J
Rosenbloom
J
J
D000595:Amino Acid Sequence; D000818:Animals; D019867:Anti-Glomerular Basement Membrane Disease / Q000276:immunology; D001323:Autoantibodies / Q000097:blood; D001324:Autoantigens / Q000737:chemistry; D001483:Base Sequence; D002417:Cattle; D003094:Collagen / Q000737:chemistry / Q000276:immunology*; D000939:Epitopes / Q000737:chemistry*; D005921:Glomerulonephritis / Q000276:immunology; D006801:Humans; D007120:Immunochemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D017433:Protein Structure, Secondary
D001323:Autoantibodies; D001324:Autoantigens; D000939:Epitopes; D010455:Peptides; D003094:Collagen