STRUCTURE OF CYTOTOXIC T LYMPHOCYTE GLYCOPROTEIN T145

细胞毒性T淋巴细胞是T淋巴细胞的亚群，其介导 生物体对病毒感染或组织的免疫反应 异体字 来自毛野豌豆种子的凝集素是 相当大的兴趣，因为一种未经表征的凝集素制剂， 据报道，其特异性结合小鼠上的糖蛋白（T145 细胞毒性T淋巴细胞 在初步工作中，三种凝集素已被 从V. villosa种子中纯化。 这些凝集素是四聚体， 由两个不同的亚基组成，具有不同的碳水化合物结合 特殊性 B4凝集素的结合，这些凝集素中的主要凝集素 种子，细胞毒性T淋巴细胞的克隆系是相当大的增加 与静息脾细胞和胸腺细胞过度结合。 在本申请中， 细胞毒性T淋巴细胞克隆系上的糖蛋白和细胞毒性T淋巴细胞克隆系上的糖蛋白。 这种糖蛋白的寡糖部分， 将详细研究V. villosa B4凝集素。 为了做到这一点，V。 将制备并表征绒毛草B4凝集素亲和吸附剂。 代谢标记的细胞毒性T淋巴细胞上的糖蛋白， 将通过SDS-PAGE和荧光法鉴定与该吸附剂结合的蛋白质。 糖肽也将从这些细胞毒性T淋巴细胞制备， 通过在V. villosa B4凝集素-琼脂糖上的亲和层析进行分级 柱 寡糖单元将从标记的 与亲和柱相互作用的糖肽和它们的主要 结构确定。 一旦这种寡糖结构被阐明， 将研究其产生的生物合成机制。 最后，将产生针对细胞毒性T细胞上的糖蛋白的抗血清。 淋巴细胞，其与V. villosa B4凝集素亲和吸附剂结合。 静息小鼠脾细胞上的免疫学相似的糖蛋白， 胸腺细胞将通过免疫沉淀溶解的 这些细胞的糖蛋白。 如果免疫学上相似的糖蛋白 存在于这些细胞上，蛋白质之间的结构相似性 糖蛋白的部分将通过胰蛋白酶肽表征 映射. 这些研究将提出一种糖蛋白 具有新的寡糖结构的蛋白质在 细胞毒性T淋巴细胞的分化，并最终导致 更好地了解细胞表面糖蛋白的功能， T淋巴细胞上的寡糖单位。

Cytotoxic T lymphocytes are a subpopulation of T lymphocytes which mediate the immune response of an organism to viral infection or tissue allographs. the lectins from the seeds of Vicia villosa are of considerable interest because an uncharacterized lectin preparation has been reported to bind specifically to a glycoprotein (T145) on mouse cytotoxic T lymphocytes. In preliminary work, three lectins have been purified from V. villosa seeds. These lectins are tetrameric and are composed of two different subunits with distinct carbohydrate binding specificities. Binding of B4 lectin, the predominant lectin in these seeds, to cloned lines of cytotoxic T lymphocytes is considerably increased over binding to resting splenocytes and thymocytes. In this application, the glycoprotein(s) on a clonel line of cytotoxic T lymphocytes and the oligosaccharide protion of this glycoprotein which specifically interacts with V. villosa B4 lectin will be studied in detail. To do this, a V. villosa B4 lectin affinity adsorbent will be prepared and characterized. The glycoproteins on metabolically-labeled cytotoxic T lymphocytes which bind to this adsorbent will be identified by SDS-PAGE and fluorography. Glycopeptides will also be prepared from these cytotoxic T lymphocytes and fractionated by affinity chromatography on the V. villosa B4 lectin-agarose column. Oligosaccharide units will be released from the labeled glycopeptides which interact with the affinity column and their primary structure determined. Once this oligosaccharide structure is elucidated, the biosythetic mechanism(s) by which it arises will be investigated. Finally, an antisera will be raised to the glycoprotein(s) on cytotoxic T lymphocytes which binds to the V. villosa B4 lectin affinity adsorbent. Immunologically similar glycoproteins on resting mouse splenocytes and thymocytes will be identified by immunoprecipitation of solubilized glycoproteins from these cells. If an immunologically similar glycoprotein is present on these cells, structural similarities between the protein portions of the glycoproteins will be characterized by tryptic peptide mapping. These studies will suggest a mechanism by which a glycoprotein with a new oligosaccharide structure is expressed during the differentiation of cytotoxic T lymphocytes and will eventually lead to a better understanding of the function of cell surface glycoproteins and their oligosacchride units on T lymphocytes.