Cross-linking of Ly 6-linked alloantigens: association between ThB and Ly 5.
The bifunctional cross-linking reagent dithiobis(succinimidyl propionate) (DSP) was used to cross-link 125I surface-labelled glycoproteins from viable thymocytes. The cells were solubilized, and the cross-linked material immunoprecipitated and analysed by SDS-PAGE. When DSP cross-linked thymocyte material was immunoprecipitated with either anti-ThB or anti-Ly 5 monoclonal antibodies, and then cleaved, molecules with masses identical to Ly 5 (Mr 180 kD) and ThB (Mr 16-18 kD) were obtained. However, if the cross-linker was not cleaved, the intact product had a molecular mass of greater than 200 kD. The identity of these co-precipitated, cross-linked moieties was formally proved by limited proteolysis peptide map analysis. The data indicated that the ThB and Ly 5 antigens were associated on the thymocyte cell surface but no such association could be found on peripheral lymphocytes. The ThB-Ly 5 interaction may indicate an association relevant to the differentiation of thymocytes.
Houlden
B A
BA
Department of Pathology, University of Melbourne, Parkville, Victoria, Australia.
McKenzie
I F
IF
Hogarth
P M
PM
D000818:Animals; D000950:Antigens, Ly / Q000032:analysis*; D000951:Antigens, Neoplasm; D000954:Antigens, Surface / Q000032:analysis*; D002478:Cells, Cultured; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D058851:GPI-Linked Proteins; D007519:Isoantigens / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D008970:Molecular Weight; D010449:Peptide Mapping; D013388:Succinimides / Q000494:pharmacology; D013601:T-Lymphocytes / Q000276:immunology*
D000950:Antigens, Ly; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D003432:Cross-Linking Reagents; D058851:GPI-Linked Proteins; D007519:Isoantigens; C035271:Ly6d protein, mouse; D008562:Membrane Glycoproteins; D013388:Succinimides; C011240:dithiobis(succinimidylpropionate)