Molecular Basis of Cadherin Mediated Cell Adhesion

钙粘蛋白介导的细胞粘附的分子基础

• 批准号: 7546548
• 负责人: LAWRENCE S SHAPIRO
• 金额: $ 30.03万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7546548
• 关键词: Adhesives; Affinity; Binding; Biological; Biological Assay; Cadherins; Cell Adhesion; Cell surface; Cell-Cell Adhesion; Defect; Development; E-Cadherin; Functional disorder; Funding; Goals; H-Cadherin; Health; Hereditary Disease; Human; Hydrogen Bonding; Individual; Knowledge; Light; Malignant Neoplasms; Maps; Mediating; Molecular; Mutagenesis; Mutation; N-Cadherin; Neoplasm Metastasis; Pattern; Pharmaceutical Preparations; Point Mutation; Positioning Attribute; Proteins; Publishing; Research; Role; Side; Site-Directed Mutagenesis; Specificity; Structure; Surface Plasmon Resonance; Testing; Vertebrates; Work; base; cell behavior; design; dimer; improved; in vitro Assay; in vivo; mutant; neoplastic cell; novel; preference; research study; small molecule; vertebrate genome

DESCRIPTION (provided by applicant): The binding of cadherins from apposing cell surfaces drives the development of organized multicellular structures. However, the physical basis for cadherin function, especially the differential binding specificities of the nineteen classical (type I and type II) cadherins, and the role of the unique GPI-anchored T-cadherin that functions in concert with them, remain poorly understood. The goals of this proposal are to provide a molecular, structural, and energetic view of classical cadherin function and to relate binding specificity at the molecular level to adhesive specificity at the cellular level. In the past funding period we made significant progress on these questions (a) by solving the first crystal structure of a complete type I cadherin ectodomain; (b) by solving the first crystal structures of type II cadherins; (c) by clearly establishing the role of 2-strand swapping in the EC1 cadherin ectodomain as the primary structural mechanism underlying classical cadherin binding specificity; (d) by developing in-vivo assays for type II cadherin function; (e) by showing that a single hydrogen bond can determine binding specificity between type I cadherins; and (f) by solving the first crystal structures of T-cadherin, showing that this non-classical cadherin functions by a novel mechanism. These advances provide much of the basis for the specific aims of the current proposal. We will (Aim 1) Determine binding affinities for an all-against-all matrix of classical cadherins and T-cadherin, and (Aim 2) employ our structural understanding of cadherins to design and characterize cadherin mutants with altered adhesive specificities. These studies will provide an improved understanding of the relationship between cadherin binding affinities and the adhesive behavior of cells, shedding new light on the functional implications of cadherin expression patterns observed in vertebrate development. Because cadherins are involved in the development of virtually all multicellular structures in vertebrate animals, their function and dysfunction have broad impact on human health. Mutations in cadherins are the underlying cause of many heritable defects. Loss of cadherin function, allowing tumor cells to de- adhere and become mobile in the body, is thought to be a prerequisite for metastasis of some forms of cancer. The research we propose will produce a comprehensive atomic-level understanding of cadherins that will be critical in understanding cadherin-related genetic disorders and can provide a basis for the development of small molecule drugs selective for inhibition of individual cadherins.

描述（由申请人提供）：钙粘蛋白与相邻细胞表面的结合驱动有组织的多细胞结构的发育。然而，钙粘蛋白功能的物理基础，特别是 19 种经典（I 型和 II 型）钙粘蛋白的差异结合特异性，以及与它们协同作用的独特的 GPI 锚定 T-钙粘蛋白的作用，仍然知之甚少。该提案的目标是提供经典钙粘蛋白功能的分子、结构和能量观点，并将分子水平的结合特异性与细胞水平的粘附特异性联系起来。在过去的资助期间，我们在这些问题上取得了重大进展：（a）通过解决完整的 I 型钙粘蛋白胞外域的第一个晶体结构； (b) 通过解析 II 型钙粘蛋白的第一个晶体结构； (c) 通过明确确定 EC1 钙粘蛋白胞外域中 2 链交换的作用作为经典钙粘蛋白结合特异性的主要结构机制； (d) 开发 II 型钙粘蛋白功能的体内测定法； (e)通过证明单个氢键可以确定I型钙粘蛋白之间的结合特异性； (f) 通过解析 T-钙粘蛋白的第一个晶体结构，表明这种非经典钙粘蛋白通过一种新机制发挥作用。这些进展为当前提案的具体目标提供了很多基础。我们将（目标 1）确定经典钙粘蛋白和 T-钙粘蛋白的所有对抗基质的结合亲和力，并（目标 2）利用我们对钙粘蛋白的结构理解来设计和表征具有改变的粘附特异性的钙粘蛋白突变体。这些研究将更好地理解钙粘蛋白结合亲和力与细胞粘附行为之间的关系，为脊椎动物发育中观察到的钙粘蛋白表达模式的功能含义提供新的线索。 由于钙粘蛋白参与脊椎动物几乎所有多细胞结构的发育，因此它们的功能和功能障碍对人类健康具有广泛的影响。钙粘蛋白突变是许多遗传缺陷的根本原因。钙粘蛋白功能的丧失使得肿瘤细胞脱离并在体内移动，这被认为是某些形式的癌症转移的先决条件。我们提出的研究将对钙粘蛋白产生全面的原子水平理解，这对于理解钙粘蛋白相关的遗传性疾病至关重要，并且可以为开发选择性抑制单个钙粘蛋白的小分子药物提供基础。