Characterization and localization of the Gova/b alloantigens to the glycosylphosphatidylinositol-anchored protein CDw109 on human platelets.
The Gova/b alloantigens are expressed on a 175-kD protein (GP175) on human platelets. Anti-Gov alloantibodies have been implicated in posttransfusion purpura and alloimmune neonatal thrombocytopenia. In this report we characterize the immunochemistry of the alloantigens and identify the platelet protein that expresses the Gov epitopes. Approximately 50% of GP175 containing the Gov epitope was released from platelets treated with phosphatidylinositol-specific phospholipase C, indicating that at least some of this protein exists as a glycosylphosphatidylinositol (GPI)-linked isoform. Radioimmunoprecipitation and immunodepletion studies indicated that the Gova/b alloantigens are expressed on the GPI-anchored CDw109 protein. The Gova/b epitopes were expressed on an extracellular, 120-kD soluble fragment (p120) of CDw109 produced by calcium-dependent protease cleavage. Anti-Gov immunoprecipitates of chymotryptic digests of p120 contained 70- and 52-kD fragments of CDw109. Deglycosylation of native CDw109 had no effect on recognition by Gov alloantisera; however, the epitopes were destroyed after exposure to sodium dodecyl sulfate. Gova/b alloantigens were expressed on platelets and PHA-activated T-cells, cultured human umbilical vein endothelial cells, and by many different tumor cell lines, consistent with the tissue distribution of CDw109.
Smith
J W
JW
McMaster University Medical Centre, Hamilton, Ontario, Canada.
Hayward
C P
CP
Horsewood
P
P
Warkentin
T E
TE
Denomme
G A
GA
Kelton
J G
JG
D000911:Antibodies, Monoclonal; D016824:Antigens, Human Platelet / Q000032:analysis*; D001792:Blood Platelets / Q000737:chemistry / Q000276:immunology*; D017261:Glycosylphosphatidylinositols / Q000276:immunology*; D006801:Humans; D007163:Immunosorbent Techniques; D010446:Peptide Fragments / Q000276:immunology; D010716:Phosphatidylinositols / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000276:immunology*; D011489:Protein Denaturation; D010738:Type C Phospholipases / Q000378:metabolism
D000911:Antibodies, Monoclonal; D016824:Antigens, Human Platelet; D017261:Glycosylphosphatidylinositols; D010446:Peptide Fragments; D010716:Phosphatidylinositols; D010980:Platelet Membrane Glycoproteins; D010738:Type C Phospholipases