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Isolation and characterization of membrane receptors for pokeweed mitogens from mouse lymphocytes

机译:小鼠淋巴细胞中商陆有丝分裂原膜受体的分离与鉴定

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pThe major glycoproteins that bind pokeweek B-cell mitogen (Pa-1) and pokeweed T-cell mitogen (Pa-2) were isolated and identified from bone-marrow-derived lymphocytes (B-cells) and thymus-derived lymphocytes (T-cells) of C3H/He mice. The surfaces of the cells were 125I-labelled by using the enzyme lactoperoxidase, and the plasma membranes were isolated from the 125I-labelled cells. These membranes were solubilized with Triton X-100 and subjected to affinity chromatography on the affinity adsorbent prepared by coupling mitogen Pa-1 or Pa-2 to activated Sepharose 4B. The glycoproteins specifically eluted with di-N-acetylchitobiose from the affinity adsorbents were analysed according to their mobility on polyacrylamide-gel electrophoresis in sodium dodecyl sulphate. These glycoproteins were further identified by immunoprecipitation with specific antisera. Immunoglobulins, possibly immunoglobulins M and D, were identified in the eluate from the B-cell membranes, but they were not detected in the eluate from the T-cell membranes. The histocompatibility-2-complex proteins (H-2D, H-2K and Ia antigens) were found to be major receptor sites for the pokeweed mitogens on both B-cells and T-cells. However, mitogen Pa-1 (B-cell) has a stronger affinity to Ia antigens than does mitogen Pa-2 (T-cell)./p
机译:>从骨髓来源的淋巴细胞(B细胞)和胸腺来源的淋巴细胞中分离并鉴定了结合p周B细胞有丝分裂原(Pa-1)和商陆T细胞有丝分裂原(Pa-2)的主要糖蛋白。 (T细胞)的C3H / He小鼠。用乳过氧化物酶对细胞表面进行125 I标记,并从125 I标记的细胞中分离出质膜。这些膜用Triton X-100溶解,并在通过将有丝分裂原Pa-1或Pa-2偶联到活化的Sepharose 4B上制备的亲和吸附剂上进行亲和层析。根据其在聚丙烯酰胺凝胶电泳中在十二烷基硫酸钠中的迁移率,分析了从亲和吸附剂中特异性用二-N-乙酰基壳二糖洗脱的糖蛋白。通过用特异性抗血清免疫沉淀进一步鉴定了这些糖蛋白。在B细胞膜的洗脱液中鉴定了免疫球蛋白,可能是免疫球蛋白M和D,但在T细胞膜的洗脱液中未检测到。发现组织相容性-2-复合蛋白(H-2D,H-2K和Ia抗原)是B细胞和T细胞上商陆有丝分裂原的主要受体位点。然而,有丝分裂原Pa-1(B细胞)对Ia抗原的亲和力比有丝分裂原Pa-2(T细胞)强。

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