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: 8006409
• 负责人: LAWRENCE S SHAPIRO
• 金额: $ 29.43万
• 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 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8006409
• 关键词: Adhesives; Affinity; Binding; Biological Assay; Cadherins; Cell Adhesion; Cell surface; Dedications; Defect; Development; Functional disorder; Funding; Goals; Grant; H-Cadherin; Health; Hereditary Disease; Human; Hydrogen Bonding; Individual; Light; Malignant Neoplasms; Mediating; Molecular; Mutation; Neoplasm Metastasis; Pattern; Pharmaceutical Preparations; Research; Role; Specificity; Structure; Vertebrates; Work; base; cell behavior; design; improved; in vivo; mutant; neoplastic cell; novel; small molecule

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）：来自贴壁细胞表面的钙粘蛋白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个经典（I型和II型）钙粘蛋白的差异结合特异性，以及独特的GPI锚定的T-钙粘蛋白与它们一起发挥作用的作用，仍然知之甚少。这个建议的目标是提供一个经典的钙粘蛋白功能的分子，结构和精力充沛的观点，并在分子水平上的结合特异性在细胞水平上的粘附特异性。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）通过解决II型钙粘蛋白的第一个晶体结构;（c）通过明确建立EC 1钙粘蛋白胞外域中的2-链交换作为经典钙粘蛋白结合特异性的主要结构机制的作用;（d）通过开发II型钙粘蛋白功能的体内测定;（e）通过显示单个氢键可以决定I型钙粘蛋白之间的结合特异性;以及（f）通过解析T-钙粘蛋白的第一晶体结构，显示这种非经典钙粘蛋白通过新机制发挥功能。这些进展为目前提案的具体目标提供了很大的基础。我们将（目的1）确定所有对所有的经典钙粘蛋白和T-钙粘蛋白矩阵的结合亲和力，并（目的2）采用我们的钙粘蛋白的结构理解，设计和表征钙粘蛋白突变体与改变的粘附特异性。这些研究将提供一个更好的理解之间的关系钙粘蛋白结合亲和力和细胞的粘附行为，脱落的新的光在脊椎动物发育中观察到的钙粘蛋白表达模式的功能意义。 由于钙粘蛋白参与脊椎动物几乎所有多细胞结构的发育，因此其功能和功能障碍对人类健康具有广泛的影响。钙粘蛋白的突变是许多遗传缺陷的根本原因。钙粘蛋白功能的丧失，允许肿瘤细胞去粘附并在体内变得移动的，被认为是某些形式的癌症转移的先决条件。我们提出的研究将产生对钙粘蛋白的全面原子水平的理解，这对于理解钙粘蛋白相关的遗传疾病至关重要，并且可以为选择性抑制单个钙粘蛋白的小分子药物的开发提供基础。