OLIGOSACCHARIDE STRUCTURE/FUNCTION IN RECOGNITION

低聚糖结构/功能的识别

• 批准号: 2748686
• 负责人: JACQUES U BAENZIGER
• 金额: $ 48.71万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-08-01 至 2004-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2748686
• 关键词: Baculoviridae; CHO cells; Sertoli cells; X ray crystallography; carbohydrate receptor; carbohydrate sequence; cell communication molecule; cell type; cysteine; gene targeting; glycoproteins; hormone regulation /control mechanism; laboratory mouse; laboratory rabbit; lectin; liver cells; luteinizing hormone; macrophage; mannose; nuclear magnetic resonance spectroscopy; oligosaccharides; receptor binding; receptor expression; surface plasmon resonance

The long term goal of this project is to determine at a molecular level how recognition of unique oligosaccharide ligands by highly specific receptors modulate both the in vivo activity of secrete glycoproteins and cellular interactions via membrane glycoproteins. We have identified a new carbohydrate binding receptor motif that recognizes oligosaccharide with the terminal sequence S04-4-GalNAcbetal, 4GlcNAcbeta1, 2Man (S4GGnM) present on a limited number of membrane and secreted glycoproteins such as the reproductive hormone LH. The S4GGnM binding site is located in the N-terminal Cysteine-Rich domain of what was considered primarily a macrophage "mannose" (Man) specific receptor that also has 8 carbohydrate recognition domains (CRD's). The Man/S4GGnM-Receptor exists in two forms that bind to either S4GGnM- or to Man-bearing ligands. Thus both the specificity and the expression of this receptor are regulated in a cell specific manner. We have shown the S4GGnM-binding form of the receptor plays a critical role in reproduction by controlling LH circuitry half life. The presence of the Man/S4ggNm-Receptor in sperm-supporting Sertoli cells, and in lymph node and spleen macrophages that are adjacent to cells expressing S4GGnM- containing ligands at their surface, suggests this receptor also plays a role in inter-cellular communication. 1) We will determine the molecular basis for differential of S4GGnM- and Man-bearing ligands by this receptor using mutagenesis in combination with structural analysis by crystallography and nuclear magnetic resonance. 2) We will determine how the expression and specificity of the Man/S4GGnM-Receptor is regulated developmentally and hormonally in and characterized to determine if their expression is coordinately regulated with that of the receptor. 3) We will generate and characterize mice that are receptor deficient and determine the impact of the biology of glycoproteins bearing these unique structures. The Man/S4GGnM-Receptor is a versatile, multi- functional receptor. Its function in any given setting will reflect the cell type expressing the receptor, the specificity of the receptor, and the character of the glycoproteins bearing oligosaccharides that are recognized. Our studies will provide new insights into the biology of this complex system that plays important roles governing cell behavior during development, reproduction, and the immune response.

这个项目的长期目标是在分子水平上确定 如何识别独特的寡糖配体的高度特异性 受体调节分泌糖蛋白的体内活性 以及通过膜糖蛋白的细胞相互作用。我们已经确定 一种新的碳水化合物结合受体基序， 具有末端序列S 04 -4-GalNAc β 1的寡糖， 4GlcNAc β 1，2 Man（S4 GGnM）存在于有限数量的膜上， 分泌的糖蛋白，如生殖激素LH。S4GGnM 结合位点位于N-末端富含半胱氨酸的结构域， 主要被认为是巨噬细胞甘露糖（Man）特异性受体 它也有8个碳水化合物识别结构域（CRD）。的 Man/S4 GGnM-受体以两种形式存在，其结合S4 GGnM-或 与人类配体的关系因此，特异性和表达 该受体以细胞特异性方式调节。我们展示了 受体的S4 GGnM结合形式在以下方面起关键作用： 通过控制LH电路半衰期来繁殖。的存在 支持精子的支持细胞和淋巴结中的Man/S4 ggNm受体 和与表达S4 GGnM的细胞相邻的脾巨噬细胞。 在其表面含有配体，表明这种受体也起着重要的作用。 在细胞间通讯中的作用。1)我们将确定 通过这种方法区分S4 GGnM和Man配体的基础 受体使用诱变结合结构分析， 晶体学和核磁共振。2)我们将决定如何 Man/S4 GGnM受体的表达和特异性受到调节， 在发育和生殖方面进行鉴定，以确定是否 它们的表达与受体的表达协调调节。第三章 我们将产生和表征受体缺陷的小鼠， 确定携带这些糖蛋白的生物学的影响， 独特的结构。Man/S4 GGnM受体是一种多功能、多功能的 功能性受体它在任何特定环境中的功能将反映 表达受体的细胞类型，受体的特异性，和 具有低聚糖的糖蛋白的特征， 认可.我们的研究将为生物学提供新的见解， 这个复杂的系统在控制细胞行为中扮演着重要的角色 在发育、繁殖和免疫反应中。