CHARACTERIZATION OF THE FC EPSILON RI ALPHA CHAIN BY CRYSTALLOGRAPHY

通过晶体学表征 FC EPSILON RI ALPHA 链

• 批准号: 3768894
• 负责人: J P KINET
• 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/3768894
• 关键词: Escherichia coli; X ray crystallography; antibody receptor; glycoprotein structure; glycosylation; immunoglobulin E; ligands; molecular site; nuclear magnetic resonance spectroscopy; receptor binding; transfection

The receptor with high affinity for IgE (FcepsilonRI) is a tetrameric complex of an IgE binding alpha chain, a beta chain and a dimer of gamma chains. In the past we have found that the extracellular region of the alpha chain is solely responsible for ligand binding. In order to further characterize the receptor binding site by crystallography, we have generated various transfectants capable of secreting large amounts of truncated alpha chain capable of binding IgE. However this material, like the native alpha is heavily glycosylated. This is preventing the formation of analyzable crystals. In order to circumvent this problem, we are following two different strategies: (1) The transfection of truncated alpha chain in cells with a defective glycosylation machinery. (2) The mutation of the seven N-linked glycosylation sites. Finally we are also attempting to produce a similar molecule in E. coli with the hope of using the NMR techniques to characterize the receptor binding site.

对IgE具有高亲和力的受体（Fc ε RI）是一种四聚体， IgE结合α链、β链和γ二聚体的复合物 店 在过去，我们已经发现，细胞外区的细胞外区域， α链仅负责配体结合。 为了 通过晶体学进一步表征受体结合位点，我们 已经产生了各种能够分泌大量 能够结合IgE的截短的α链。 然而，这种材料， 就像天然的α是高度糖基化的。 这是为了防止 形成可分析的晶体。 为了规避这个问题， 我们遵循两种不同的策略：（1）转染 在糖基化机制有缺陷的细胞中截短的α链。 (2)7个N-连接糖基化位点的突变。 最后我们 也试图在E.大肠杆菌， 希望使用核磁共振技术来表征受体结合 绝佳的价钱