STRUCTURE AND FUNCTION OF THE HIGH AFFINITY IGE RECEPTOR

高亲和力 IGE 受体的结构和功能

• 批准号: 2071665
• 负责人: MICHAEL W ROBERTSON
• 金额: $ 11.9万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2071665
• 关键词: antibody receptor; chimeric proteins; immunoglobulin E; molecular cloning; nuclear magnetic resonance spectroscopy; point mutation; protein engineering; protein structure function; receptor binding

The high-affinity receptor (Fc epsilon RI) specific for immunoglobulin E (IgE) is found primarily on mast cells and basophils. Activation of these cells occurs in response to crosslinking of Fc epsilon RI with IgE-antigen complexes which initiates a cascade of cellular events culminating in the release of pharmacologically active substances that contribute to the clinical symptoms of many atopic disorders such as allergy, allergic asthma and immediate hypersensitivity. In this research program the structural features of Fc epsilon RI that contribute to specific receptor function will be explored. One goal is to define the molecular features of the Fc epsilon RI alpha-subunit involved in IgE binding. This will be approached using the dual approach of (i) protein engineering studies using both domain and point mutagenesis procedures and (ii) structure elucidation studies to be conducted initially using nuclear magnetic resonance spectroscopy in a collaborative study with Dr. A. Bax, National Institutes of Health. A parallel, but longer-term goal, is to generate receptor fragment crystals suitable for eventual crystallographic studies. Protein engineering studies will extensively utilize the powerful technique of phage display together with bacterial expression of Fc epsilon RI alpha fragments with subsequent physical characterization of ligand-binding activity. A long-term goal of this study is the precise definition of the 3-dimensional configuration of a "minimum complex" of Fc epsilon RI alpha-IgE. An additional but lower priority aim of this Proposal is to characterize the rat alpha-subunit ectodomain through detailed protein engineering studies, with the eventual goal of elucidating the 3-dimensional structure of this IgE-binding molecule. The requirement for Fc epsilon RI beta and/or gamma-subunit coexpression for rat-human alpha-chain ectodomain chimera will be explored as a means of probing the presumptive extracellular interaction of the receptor subunits. Progress in all of these studies should assist in the rational design of potential therapeutics for the treatment of atopic disorders. In addition, results should facilitate molecular studies of other immunologically important receptors [such as Fc gamma R] which are expected to share significant structural features with the IgE mast cell receptor.

免疫球蛋白E的高亲和力受体(Fc Epsilon RI) (IgE)主要存在于肥大细胞和嗜碱性粒细胞上。激活这些 细胞对Fc epsilon RI与IgE抗原的交联反应 引发一系列细胞事件的复合体，最终导致 释放的药理活性物质有助于 许多特应性疾病的临床症状，如过敏、过敏 哮喘和即刻过敏。在这个研究项目中， Fc epsilon RI参与特异性受体的结构特征 我们将探索它的功能。一个目标是定义分子特征 参与IgE结合的Fc epsilon RIα亚基。这将是 用对偶方法探讨(I)蛋白质工程研究 同时使用结构域和点突变程序和(Ii)结构 初步使用核磁进行的澄清研究 与美国国立卫生研究院A.Bax博士的合作研究中的共振光谱 卫生研究院。一个平行但更长期的目标是，产生 适合于最终结晶学研究的受体片段晶体。 蛋白质工程研究将广泛利用强大的 Fc噬菌体展示及细菌表达技术 Epsilon RIα片段以及随后的物理特征 配体结合活性。这项研究的长期目标是准确地 Fc的“最小复形”的三维构型定义 Epsilon RIα-IgE。这是一个额外但优先级较低的目标 建议通过以下方式来表征RATα亚基胞外结构域 详细的蛋白质工程研究，最终目标是 阐明了这种IgE结合分子的三维结构。这个 Fc epsilon RIβ和/或伽马亚基共表达的要求 大鼠-人α链胞外结构域嵌合体将作为一种手段进行探索 探讨受体的胞外相互作用 亚单位。所有这些研究的进展应该有助于理性的 治疗特应性疾病的潜在疗法的设计。 此外，这些结果应该有助于其他生物的分子研究。 免疫上重要的受体[如Fc-Gamma R] 预计将与IgE肥大细胞共享显著的结构特征 受体。