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CROSS‐LINKING OF Ly 6‐LINKED ALLOANTIGENS: ASSOCIATION BETWEEN ThB AND Ly 5
Author(s) -
Houlden B. A.,
McKenzie I. F. C.,
Hogarth P. M.
Publication year - 1989
Publication title -
international journal of immunogenetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.41
H-Index - 47
eISSN - 1744-313X
pISSN - 1744-3121
DOI - 10.1111/j.1744-313x.1989.tb00445.x
Subject(s) - thymocyte , immunoprecipitation , monoclonal antibody , glycoprotein , bifunctional , chemistry , microbiology and biotechnology , proteolysis , propionate , polyclonal antibodies , biology , biochemistry , antigen , antibody , immunology , cd8 , enzyme , gene , catalysis
SUMMARY The bifunctional cross‐linking reagent dithiobis (succinimidyl propionate) (DSP) was used to cross‐link 125 I 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 ( M r 180 kD) and ThB ( M r 16–18 kD) were obtained. However, if the cross‐linker was not cleaved, the intact product had a molecular mass of > 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.