Identification of a surface protein (p100) associated with two glycosyl-phosphatidylinositol-linked molecules (Thy-1 and ThB) by natural anti-lymphocyte autoantibodies.
Our previous study of natural autoantibodies showed that anti-lymphocyte antibodies are frequently produced by perinatal B cells from normal strains of mice. One-third of these monoclonal antibodies (mAb) recognized similar epitopes on the surface of thymocytes. In the present report, we have characterized the molecule recognized by three of these mAb (D10, G7, 22). These mAb identified a 100-kDa protein (p100) on the surface of thymocytes. This protein resolved into 70-kDa polypeptide chains under reducing conditions. Inhibition experiments as well as antibody immunoprecipitations in the presence of mild detergents revealed non-covalent association of the p100 with Thy-1 and ThB. A similar multimolecular complex was identified following chemical cross-linking of thymocyte surface proteins. Analysis of several Thy-1-defective mutant cells lines, and thymocytes treated with phosphatidylinositol-specific phospholipase C (PI-PLC) showed that the expression of p100 was strongly influenced by Thy-1 molecule. The p100 was resistant to PI-PLC treatment and was not released into the supernatant as was the case for Thy-1 and ThB molecules. These data lead us to propose that the p100 is a transmembrane protein, the expression of which in the plasma membrane is dependent on the association or presence of Thy-1 molecule.
Department of Microbiology, University of Alabama, Birmingham.
A
A
Lehuen
R C
RC
Monteiro
J F
JF
Kearney
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis*; D001323:Autoantibodies / Q000276:immunology*; D002793:Cholic Acids / Q000494:pharmacology; D005260:Female; D006017:Glycolipids / Q000032:analysis*; D017261:Glycosylphosphatidylinositols; D008214:Lymphocytes / Q000276:immunology*; D008297:Male; D008562:Membrane Glycoproteins / Q000032:analysis*; D008565:Membrane Proteins / Q000032:analysis*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D010716:Phosphatidylinositols / Q000032:analysis*; D018800:Thy-1 Antigens; D010738:Type C Phospholipases / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D001323:Autoantibodies; D002793:Cholic Acids; D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D010716:Phosphatidylinositols; D018800:Thy-1 Antigens; D010738:Type C Phospholipases; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate