IMMUNOPHILIN AND RELATED PROTEINS

最近发现了免疫抑制药物的结合蛋白 与胞质类固醇的低聚体结构有关 感受器。结合蛋白与已知的 免疫抑制药物，如雷帕霉素、FK5O6和环孢素A和 因为免疫系统的功能有升有降 随着系统老化等情况的出现，似乎存在 一种或多种内源性免疫抑制剂，其水平可能低于 各种类型的控制。在研究的过程中， 糖皮质激素受体非DNA结合复合体的纯化 蛋白质，我们发现了一种新的与FKBP-52(p55-59)络合的蛋白质。 免疫亲和素。因为这种新的蛋白质是用FKBP-52从 不同的细胞系，只有当FKBP-52出现时，它才会出现，我们得出结论 它可能在细胞周期的信号转导途径中发挥作用。 免疫抑制作用。因此，我们计划进行以下工作 目的：(1)FKBP-52全长、截短和突变 在杆状病毒系统中过度表达。定点突变将是 然后是突变蛋白的过度表达和所有这些 蛋白质将被提纯。(2)高效表达和纯化FKBP-52、hsp90、 HSP7O和其他胞浆因子将被重组为高 分子量复合体，与细胞复合体相当，以及 免疫抑制药物配体对这些复合体的影响将是 评估过了。(3)与59 kDa FKBP-52相关的新的磷酸蛋白将是 克隆、过表达、纯化和鉴定。(4)中的序列 FKBP-52负责结合hsp90、hsp70、ATP和 59 kDa FKBP-52相关蛋白将通过突变确定 FKBP-52的cDNA，过表达衍生蛋白并组装成 高分子量的复合体。(5)针对FKBP的特异性抗体- 52及其相关的磷蛋白将被开发出来。这些实验 将阐明信号转导过程中涉及的途径 药物诱导的免疫抑制，可能有助于所需的方法 发现内源性免疫抑制药，结果将直接 影响器官和组织移植。

Binding proteins for immunosuppressant drugs have been discovered recently in connection with the oligomeric structures of cytoplasmic steroid receptors. The binding proteins form complexes with known immunosuppressant drugs, such as rapamycin, FK5O6 and cyclosporin A and because there are elevations and declines in the functioning of the immune system with aging and other conditions, it seems likely that there exist one or more endogenous immunosuppressant agents whose levels may be under various types of control. In studying the reassembly of the glucocorticoid receptor non-DNA-binding complex form with purified proteins, we discovered a new protein complexed with the FKBP-52 (p55-59) immunophilin. Because this new protein is purified with the FKBP-52 from different cell lines and it appears only when FKBP-52 appears, we conclude that it may function in the signal transduction pathway of immunosuppressant action. Consequently, we plan to pursue the following objectives: (1) Full-length, truncated and mutated FKBP-52 will be overexpressed in the baculovirus system. Site directed mutagenesis will be followed by overexpression of the mutant proteins and all of these proteins will be purified. (2) Overexpressed and purified FKBP-52, hsp90, hsp7O and other cytosolic factors will be reconstituted into high molecular weight complexes, comparable to cellular complexes, and the effects of immunosuppressant drug ligands on these complexes will be assessed. (3) The 59kDa FKBP-52-associated new phosphoprotein will be cloned, overexpressed, purified and characterized. (4) The sequences in FKBP-52 that are responsible for binding to hsp9O, hsp7O, ATP and the 59kDa FKBP-52-associated protein will be determined by mutation of the FKBP-52 cDNA, overexpression of the derivative protein and assembly into the high molecular weight complex. (5) Specific antibodies against FKBP- 52 and its associated phosphoprotein will be developed. These experiments will clarify the signal transduction process involved in the pathway of drug-induced immunosuppression and may contribute to the methods required to discover the endogenous immunosuppressants, results that will directly affect organ and tissue transplantation.