ZAP-70, A T CELL RECEPTOR ZETA CHAIN ASSOCIATED PROTEIN

自身免疫性疾病和风湿性疾病的谱跨越多种类型， 免疫反应的缺陷然而，要理解潜在的 这些疾病的病理生理学，从而能够开发治疗方法 在一个合理的方式，免疫反应的生化基础必须 被理解T细胞是主要的效应细胞之一， 免疫系统的调节器。它在宿主防御中的作用包括杀死 病毒感染细胞（细胞毒性），其他细胞的免疫调节 参与免疫应答（例如，B细胞-T细胞相互作用），和 分泌淋巴因子以影响多种组织的能力， 细胞外源病原体激活T细胞导致这些 效应子功能由T细胞抗原受体（TCR）介导。超过 在过去的十年中，TCR已经被生物化学定义，其结构 对正常装配和功能的要求进行了详细研究。 然而，TCR介导的信号被传递到 效应器功能仍然不清楚。多种细胞内 生物化学信号已经被牵连，包括蛋白质 通过酪氨酸和丝氨酸/苏氨酸蛋白激酶的磷酸化， 磷酸酶的去磷酸化和磷酸肌醇的移动 导致细胞质钙水平增加。但 这些生物化学过程的相互作用尚不清楚。 本提案的目的是剖析和定义信号级联 参与TCR活化。欧文和韦斯最近证明了 Z-链胞质结构域在TCR信号中的重要性 转导此外，我们最近还描述了一种蛋白质，ZAP- 70，与zeta相关，并在 刺激TCR。本建议采用以下组合 生物化学、分子和免疫学方法来表征ZAP-70。 具体而言，这些措施包括：l。ZAP-70的蛋白纯化 微测序，2.利用分离编码ZAP-70的cDNA克隆 由蛋白质序列产生的简并寡核苷酸，和3. 产生针对ZAP-70的抗体。总之，推导出的序列 ZAP-70可以提供基于序列同源性的对其功能的洞察。 此外，这些结构研究和试剂的可用性， ZAP-70将允许进一步的生物学和功能研究。这些 研究将使我们更好地了解T细胞的激活， 外来病原体的细胞。

The spectrum of autoimmune and rheumatic diseases spans a variety of defects in the immune response. However, to appreciate the underlying pathophysiology of these diseases and thus be able to develop therapeutics in a rational fashion, the biochemical basis of the immune response must be understood. The T cell represents one of the principal effectors and regulators of the immune system. Its role in host defense includes killing of virally infected cells (cytotoxicity), immunoregulation of other cells involved in the immune response (e.g., B cell-T cell interactions), and the ability to secrete lymphokines to effect a variety of tissues and cells. Activation of T cells by foreign pathogens resulting in these effector functions is mediated by the T cell antigen receptor (TCR). Over the past decade, the TCR has been defined biochemically and its structural requirements for normal assembly and function studied in great detail. However, the mechanism by which TCR-mediated signals are transmitted into effector functions remain poorly defined. A variety of intracellular biochemical signals have been implicated, including protein phosphorylation via both tyrosine and serine/threonine protein kinases, dephosphorylation by phosphatases, and mobilization of inositol phosphate with resultant increases in cytoplasmic calcium levels. However, the interplay of these biochemical processes are not understood. The aim of this proposal is to dissect and define the cascade of signals involved in TCR activation. Irving and Weiss have recently demonstrated the importance of the cytoplasmic domain of the zeta-chain in TCR signal transduction. Furthermore, we have also recently described a protein, ZAP- 70, that associates with zeta and undergoes tyrosine phosphorylation after stimulation of the TCR. This proposal utilizes a combination of biochemical, molecular, and immunologic approaches to characterize ZAP-70. Specifically, these include: l. purification of ZAP-70 for protein microsequencing, 2. isolation of cDNA clones encoding for ZAP-70 utilizing degenerate oligonucleotides generated from protein sequence, and 3. generation of antibodies against ZAP-70. Together, the deduced sequence of ZAP-70 may provide insight into its function based on sequence homology. Moreover, these structural studies and availability of reagents against ZAP-70 will permit further biologic and functional investigation. These studies will allow for a greater understanding of the activation of T cells by foreign pathogens.