Characterization of human lymphocyte surface receptors for mitogenic and non-mitogenic substances.
To compare the receptor patterns for mitogenic and non-mitogenic substances, surface glycoproteins of human lymphocytes were labelled with the lactoperioxidase-catalysed iodination technique and with a galactose oxidase-tritiated sodium borohydride technique. Labelled cells were detergent-solubilized, and the lysates were allowed to react with insolubilized purified mitogenic lectins, phytohaemagglutinin, leucoagglutinin and an insolubilized non-mitogenic lectin, oxidized leucoagglutinin. Lectin-reactive proteins were eluted with sodium dodecyl sulphate (SDS) buffer. Cell membrane components reactive with anti-lymphocyte globulin (ALG) were retrieved by indirect immunoprecipitation with protein-A-bearing staphylococcus Cowan I strain (SaCI). Lectin- and ALG-reactive proteins were analysed by SDS polyacrylamide gel electrophoresis. Iodinated glycoproteins regularly showed four major components with molecular weights of 120,000, 70,000, 60,000 and 43,000 daltons, respectively, on 7% gels. An additional broad peak in the molecular weight range 20,000--35,000 daltons was found on 10% gels. Tritiated glycoproteins also showed four major components with MW 120,000, 70,000, 60,000 AND 42,000, RESPECTIVely, which reacted with lectin and ALG. In addition, ALG reacted with some glycoproteins with MW between 150,000 and 230,000 daltons. On 10% gels additional lectin- and ALG-binding glycoproteins with MW around 30,000 daltons were found. The similarity in structures bound by mitogenic and non-mitogenic substances indicates that lymphocyte activation may depend on some property conferred by the mitogen.
D000961:Antilymphocyte Serum / Q000494:pharmacology; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis / Q000276:immunology*; D006801:Humans; D007553:Isotope Labeling; D008214:Lymphocytes / Q000276:immunology*; D008934:Mitogens / Q000494:pharmacology; D008970:Molecular Weight; D010835:Phytohemagglutinins / Q000494:pharmacology; D011975:Receptors, Mitogen / Q000276:immunology*
D000961:Antilymphocyte Serum; D006023:Glycoproteins; D008934:Mitogens; D010835:Phytohemagglutinins; D011975:Receptors, Mitogen