Anti-mitochondrial flavoprotein autoantibodies of patients with myocarditis and dilated cardiomyopathy (anti-M7): interaction with flavin-carrying proteins, effect of vitamin B2 and epitope mapping.
Vitamin B2 and flavin cofactors are transported tightly bound to immunoglobulin in human serum. We reasoned that anti-mitochondrial flavoprotein autoantibodies (alpha Fp-AB) present in the serum of patients with myocarditis and cardiomyopathy of unknown aetiology may form immunoglobulin aggregates with these serum proteins. However, immunodiffusion and Western blot assays demonstrated that the flavin-carrying proteins were not recognized by alpha Fp-AB. Apparently the flavin moiety in the native protein conformation was inaccessible to alpha Fp-AB. This conclusion was supported by the absence of an immunoreaction between the riboflavin-binding protein from egg white and alpha FP-AB. Intravenous application of vitamin B2 to rabbits immunized with 6-hydroxy-D-nicotine oxidase, a bacterial protein carrying covalently attached FAD, did not neutralize alpha Fp-AB which had been raised in the serum of the animals. FAD-carrying peptides generated from 6-hydroxy-D-nicotine oxidase by trypsin and chymotrypsin treatment were not recognized by the alpha Fp-AB, but those generated by endopeptidase Lys were. This demonstrates that the epitope recognized by alpha Fp-AB comprises, besides the flavin moiety, protein secondary structure elements.
Stähle
I
I
Institute of Biochemistry and Molecular Biology, University of Freiburg, Germany.
Brizzio
C
C
Barile
M
M
Brandsch
R
R
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D000937:Antigen-Antibody Reactions / Q000187:drug effects; D001323:Autoantibodies / Q000097:blood*; D001324:Autoantigens / Q000235:genetics; D002311:Cardiomyopathy, Dilated / Q000276:immunology*; D002352:Carrier Proteins / Q000276:immunology; D016022:Case-Control Studies; D018604:Epitope Mapping; D005420:Flavoproteins / Q000235:genetics / Q000276:immunology*; D006801:Humans; D026901:Membrane Transport Proteins; D008928:Mitochondria / Q000276:immunology; D008969:Molecular Sequence Data; D009205:Myocarditis / Q000276:immunology*; D009206:Myocardium / Q000276:immunology; D009500:Neutralization Tests; D011817:Rabbits; D051381:Rats; D012256:Riboflavin / Q000378:metabolism / Q000494:pharmacology
D001323:Autoantibodies; D001324:Autoantigens; D002352:Carrier Proteins; D005420:Flavoproteins; D026901:Membrane Transport Proteins; C019079:riboflavin-binding protein; D012256:Riboflavin