Salisbury J G, Banga J P
Thymus. 1985;7(6):335-44.
The surface glycoproteins of rat large, mainly dividing, thymocytes and their small, non-dividing, progeny have been examined by labelling with 3H-sodium borohydride. A prominent glycoprotein component of apparent mol. wt. 150 K was labelled on both the cell types and by immunoprecipitation with a specific monoclonal antibody, was demonstrated to contain the leucocyte common antigen (LCA). The binding of this glycoprotein to Lens culinaris lectin (LcH) has been studied. Preliminary results obtained by different binding of ratiolabelled LcH to sodium dodecyl sulphate (SDS) solubilised crude plasma membrane from large and small thymocytes, fractionated by polyacrylamide gel electrophoresis (PAGE) indicated that a 150 K apparent molecular weight glycoprotein in the small cells bound radiolabelled LcH whereas virtually no binding of the lectin was observed in the 150 K region for large cells. However, by utilizing a different approach of lectin affinity chromatography on LcH-Sepharose 4B of solubilized NaB3H4 labelled membrane, followed by immunoprecipitation, we unequivocally showed that the 150 K labelled glycoprotein of both large and small cells was bound to LcH lectin and only eluted with the specific sugar alpha-methyl pyranoside (alpha-MP). The implications of these results is discussed.
通过用³H-硼氢化钠标记，对大鼠主要处于分裂期的大型胸腺细胞及其处于非分裂期的小型子代细胞的表面糖蛋白进行了检测。两种细胞类型均标记上了一种表观分子量为150K的显著糖蛋白成分，通过用一种特异性单克隆抗体进行免疫沉淀，证明其含有白细胞共同抗原（LCA）。对这种糖蛋白与菜豆凝集素（LcH）的结合进行了研究。通过将放射性标记的LcH与经十二烷基硫酸钠（SDS）溶解的来自大型和小型胸腺细胞的粗质膜进行不同比例结合，再经聚丙烯酰胺凝胶电泳（PAGE）分离后获得的初步结果表明，小型细胞中一种表观分子量为150K的糖蛋白结合了放射性标记的LcH，而在大型细胞的150K区域几乎未观察到凝集素的结合。然而，通过采用一种不同的方法，即对经硼氢化钠标记的溶解膜在LcH-琼脂糖4B上进行凝集素亲和层析，随后进行免疫沉淀，我们明确表明，大型和小型细胞的150K标记糖蛋白均与LcH凝集素结合，且仅用特异性糖α-甲基吡喃糖苷（α-MP）洗脱。讨论了这些结果的意义。
