BINDING OF PEPTIDES TO MHC MOLECULES

肽与 MHC 分子的结合

• 批准号: 6160663
• 负责人: D H MARGULIES
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6160663
• 关键词: CD8 molecule; MHC class I antigen; T cell receptor; T lymphocyte; antigen presentation; laboratory mouse; molecular cloning; tissue /cell culture

The objective of this work has been to understand the details of the interaction of self and foreign peptides with MHC molecules, as studied both by in vitro binding methods and by assays of T cell activation. As this project has developed, we have developed a system in which the binding of the antigenic peptide to the MHC class I molecule H-2Ld is measured and in which we can in parallel measure the interaction of the MHC/peptide complex with the T cell receptor. Current experiments have investigated the influence of the coreceptor molecule, CD8, on the MHC/TCR interaction as well as the nature of the early signaling events that are elicited by the stimulation of the T cell with peptide/MHC complexes and those containing variant peptides. Attention has been focused on a paradoxical result: the identification of a single peptide variant that complexes with the mouse MHC class I molecule H-2Ld, serves as an effective sensitizing peptide, but fails to form a complex in which we can detect binding by surface plasmon resonance. The variant peptide seems to form a complex that activates the T cells to cytolysis, lymphokine production, or now, as studied by Dr. M. Jelonek in collaboration with Dr. I. Stefanova by early phosphorylation events. Since there has been such a discrepancy in the ability of the variant peptide/H-2Ld complex to bind the TCR in vitro it is remarkable that the T cell response seems to show only minor kinetic and quantitative differences in early phosphorylation patterns. Attempts to evaluate the contribution of CD8 (produced by Australian collaborators Brendan Classon and Peter Hudson) to the binding reaction of H-2Ld/peptide complexes with the cognate TCR revealed interaction of the CD8 to H-2Ld molecules devoid of peptide. This suggested that an H-2Ld peptide motif in the amino terminal region of the CD8 protein was mediating binding to the MHC molecule through the MHC's peptide binding groove. A variety of direct binding and competition experiments have confirmed that H-2Ld can carry out this mode of binding to the mature CD8.

这项工作的目的是了解 研究表明，自身肽和外源肽与 MHC 分子的相互作用 通过体外结合方法和 T 细胞活化测定。 随着这个项目的发展，我们开发了一个系统，其中 抗原肽与 MHC I 类分子 H-2Ld 的结合是 测量并且我们可以并行测量 MHC/肽与 T 细胞受体的复合物。 目前的实验有 研究了辅助受体分子 CD8 对 MHC/TCR 相互作用以及早期信号事件的性质 由肽/MHC 刺激 T 细胞引起 复合物和含有变异肽的复合物。 人们的注意力集中在一个矛盾的结果上： 与小鼠 MHC I 类复合的单一肽变体 分子 H-2Ld，作为有效的致敏肽，但失败了 形成复合物，我们可以在其中检测表面等离子体的结合 谐振。 变异肽似乎形成了一种复合物，可以激活 T 细胞进行细胞溶解、淋巴因子产生，或者现在，正如研究 M. Jelonek 博士与 I. Stefanova 博士早期合作 磷酸化事件。 既然出现了这样的差异 变体肽/H-2Ld复合物在体外结合TCR的能力 值得注意的是，T 细胞反应似乎只表现出轻微的 早期磷酸化模式的动力学和定量差异。 尝试评估CD8（澳大利亚制作）的贡献 合作者布伦丹·克拉森（Brendan Classon）和彼得·哈德森（Peter Hudson）对结合反应进行了研究 H-2Ld/肽复合物与同源 TCR 的相互作用揭示了 CD8 为不含肽的 H-2Ld 分子。 这表明 CD8 蛋白氨基末端区域的 H-2Ld 肽基序为 通过 MHC 肽结合介导与 MHC 分子的结合 槽。 各种直接结合和竞争实验 证实H-2Ld可以进行这种与成熟细胞结合的模式 CD8。