Immunochemical characterization of an autoantigen on platelet glycoprotein IIb in chronic ITP: comparison with the Baka alloantigen.
Employing an immunoblotting procedure, we have identified and characterized an autoantigen carried on glycoprotein (GP) IIb in a patient with chronic idiopathic thrombocytopenic purpura (ITP), and have compared the location of the autoantigen with that of the platelet-specific alloantigen Baka. Immunoblots, using the partially purified GP IIb/IIIa complex as the target antigen, indicated that GP IIb alpha carried both the ITP autoantigen and the Baka alloantigen. The ITP plasma contained another antibody against a 100 kD protein (P100), a trace contaminant in the GP IIb/IIIa sample, which is probably a proteolytic fragment of an internal 124 kD protein. After chymotrypsin treatment, the auto- and alloantigen were found to be located on 65 kD fragments detectable under reducing conditions. In addition, immunoblots made after two-dimensional nonreduced-reduced SDS-polyacrylamide gel electrophoresis (SDS-PAGE) directly demonstrated that both 65 kD fragments had a molecular weight of 80 kD under nonreducing conditions; this provides evidence that these fragments were one and the same, and were derived from GP IIb alpha. Immunoblots of platelets digested in situ with chymotrypsin indicated that the 65 kD fragment of GP IIb alpha was retained by the platelet membrane. We conclude, therefore, that a 65 kD fragment, which represents the membrane side of the chymotrypsin cleavage site on GP IIb alpha, carries a clinically important determinant(s) recognized not only by the anti-Baka alloantibody, but also by the ITP autoantibody.
Tomiyama
Y
Y
Second Department of Internal Medicine, Osaka University Medical School, Japan.
Kurata
Y
Y
Shibata
Y
Y
Honda
S
S
Furubayashi
T
T
Mizutani
H
H
Tsubakio
T
T
Yonezawa
T
T
Tarui
S
S
D000328:Adult; D016824:Antigens, Human Platelet; D001324:Autoantigens / Q000032:analysis*; D001792:Blood Platelets / Q000276:immunology; D000939:Epitopes / Q000032:analysis; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007518:Isoantibodies / Q000276:immunology; D007519:Isoantigens / Q000032:analysis*; D010446:Peptide Fragments / Q000276:immunology; D010980:Platelet Membrane Glycoproteins / Q000276:immunology*; D011696:Purpura, Thrombocytopenic / Q000097:blood / Q000276:immunology*
C044561:3a alloantigen, human; D016824:Antigens, Human Platelet; D001324:Autoantigens; D000939:Epitopes; D007518:Isoantibodies; D007519:Isoantigens; D010446:Peptide Fragments; D010980:Platelet Membrane Glycoproteins