Electrophoretic studies of antimitochondrial antibodies: identification of mitochondrial antigens specific to primary biliary cirrhosis.
Mitochondrial inner membrane proteins extracted from beef heart tissue were examined for reactivity to antimitochondrial antibody (AMA) present in sera of patients with primary biliary cirrhosis (PBC) by an immunoblotting technique. Four proteins, which reacted with AMA, had molecular masses of 70 kDa, 50 kDa, 47 kDa and 40 kDa, as defined by their relative mobility (Rf) in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. All sera of 114 PBC patients were positive with at least one and as many as four of the mitochondrial proteins. The major antigenic proteins of mitochondrial inner membrane to which AMA reacts were the 70 kDa and 47 kDa proteins. All PBC sera containing antibodies to the 50 kDa and/or 40 kDa proteins reacted with 70 kDa as well. The isolation of antigen reacting with AMA of PBC is important to warrant further study of AMA and the cause of the disease. The isolation of responsible antigens had been difficult because the four antigens were insoluble. However, the antigen newly found by us, the 36 kDa fragment, obtained by partial trypsin digestion, is soluble. Using several procedures, the antigenic protein target of AMA was purified from mitochondria for the first time. We determined the N-terminal sequence of the soluble 36 kDa fragment, 25 residues in length. Until now the N-terminal sequence of the 36 kDa protein has not shown significant homology with any known protein. The present results of antigen purification would contribute to the elucidation of the epitopes of AMA antigen.
Ishii
H
H
Department of Gastroenterology, Juntendo University School of Medicine, Tokyo, Japan.
Muno
D
D
Namihisa
T
T
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000941:Antigens / Q000032:analysis* / Q000302:isolation & purification; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005260:Female; D007525:Isoelectric Focusing / Q000379:methods; D008105:Liver Cirrhosis, Biliary / Q000276:immunology*; D008297:Male; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D008928:Mitochondria / Q000201:enzymology / Q000276:immunology*; D008970:Molecular Weight; D009206:Myocardium / Q000032:analysis; D010446:Peptide Fragments / Q000032:analysis; D051381:Rats; D014357:Trypsin
D000596:Amino Acids; D000906:Antibodies; D000941:Antigens; D008565:Membrane Proteins; D010446:Peptide Fragments; D014357:Trypsin; D000251:Adenosine Triphosphatases