INTERCADHERIN INTERACTIONS IN DESMOSOMES

桥粒中钙粘蛋白间的相互作用

• 批准号: 7483747
• 负责人: Sergey M Troyanovsky
• 金额: $ 19.09万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7483747
• 关键词: Address; Adherens Junction; Adhesions; Adhesives; Appendix; Autoantibodies; Autoimmune Process; Biological Assay; Biological Models; Bulla; Cadherins; Cell Adhesion Molecules; Cell membrane; Cell-Cell Adhesion; Cells; Chemistry; Co-Immunoprecipitations; Complex; Cysteine; Cytoskeleton; Desmosomes; Development; Dimerization; Disease; Drug Delivery Systems; Endocytosis; Epidermis; Epithelial; Epithelial Cells; Event; Face; Facility Construction Funding Category; Foundations; Future; Glues; Grant; Human; In Vitro; Knowledge; Laboratories; Lateral; Life; Link; Mechanics; Mediating; Modeling; Molecular; Monitor; Neoplasm Metastasis; Numbers; Object Attachment; Pemphigus; Pemphigus Vulgaris; Pharmaceutical Preparations; Pharmacy (field); Process; Property; Proteins; Resistance; Signal Transduction; Site; Skin; Skin Abnormalities; Solid; Specific qualifier value; Stress; Structure; System; Tissues; Vaccines; Work; Wound Healing; adhesion receptor; cadherin 15; cohesion; crosslink; dimer; extracellular; fibrosarcoma; improved; interest; keratinocyte; mutant; protein protein interaction; receptor; reconstitution; research study; scaffold; segregation; skin disorder; tumor growth

DESCRIPTION (provided by applicant): Classic and desmosomal cadherins are ubiquitous transmembrane adhesion receptors of epithelial cells. While significant progress has been made toward understanding classic cadherin adhesion, the mechanisms of adhesion by desmosomal cadherins have been poorly studied. Two factors - the complexity of desmosome cadherin composition and the insolubility of desmosomes - have hampered direct examination of protein-protein interactions in these structures. However, significant progress in understanding classic cadherin adhesion as well new assays that we developed have enabled us to address the question of cadherin-cadherin interactions in desmosomes on a more sophisticated level. The key Specific Aim of this application is the development of a site-specific cross-linking assay that is able to monitor intercadherin interactions in desmosomes. Such an assay is essential for clarifying many open questions about the mechanisms of desmosome formation. Among them are the following: (i) whether desmosomal cadherins are able to produce strand dimers; (ii) whether these dimers are heterodimers, homodimers, or both; (iii) whether this dimerization process can be reproduced in vitro or if it can proceed only in living cells within desmosomes; (iv) whether intracellular desmosomal proteins regulate extracellular adhesive interactions; and (v) whether desmosomes have a specific lateral alignment of desmosomal cadherins. Our previous experiments with classic cadherins provide a clear blueprint for such a study and indicate that this project is feasible. This application outlines a project to resolve a very difficult-to-study but critical aspect of cell-cell adhesion - the protein-protein adhesive interactions in desmosomes. Desmosomes are major adhesion structures in the epidermis that glue together keratinocytes. Despite their significance, it is not known how desmosomes form the tight, ordered and dynamic adhesive links between epidermal cells. Understanding this question is necessary for elucidating the mechanisms of severe human blistering skin diseases, including pemphigus foliaceus and pemphigus vulgaris. Furthermore, such knowledge will specify targets for compounds that decrease cohesion between epidermal cells. These compounds may have applications in wound healing and in promoting fast delivery of other drugs or vaccines throughout the skin. Since desmosomes are involved in intracellular signaling, the manipulation of their adhesive interactions may present a new avenue for modulating intracellular signals in order to correct hyperproliferation, abnormal differentiation and other skin abnormalities.

描述（由申请人提供）：经典和脱染色体钙粘着蛋白是上皮细胞无处不在的跨膜粘附受体。虽然在理解经典的钙粘蛋白粘附方面取得了重大进展，但对脱乳小蛋白的粘附机制的研究很差。两个因素 - 脱粘性钙粘蛋白组成的复杂性和脱糖体的不溶性 - 阻碍了对这些结构中蛋白质 - 蛋白质相互作用的直接检查。但是，在理解经典的钙粘蛋白粘附以及我们开发的新测定方面的重大进展使我们能够在更复杂的水平上解决脱糖体中钙粘蛋白 - 钙粘蛋白相互作用的问题。该应用程序的主要特定目的是开发特定于位点特定的交联测定法，该测定能够监测脱糖体中的钙粘蛋白相互作用。这样的测定对于阐明许多关于脱糖体形成机制的开放问题至关重要。其中包括以下内容：（i）脱乳小体钙粘蛋白是否能够产生链二聚体； （ii）这些二聚体是异二聚体，同型二聚体还是两者兼而有之； （iii）该二聚过程是否可以在体外再现，还是只能在脱糖体内的活细胞中进行； （iv）细胞内脱发蛋白是否调节细胞外粘合剂相互作用； （v）脱糖体是否具有特异性的脱骨钙蛋白的侧向比对。我们以前使用经典钙粘蛋白的实验为这项研究提供了明确的蓝图，并表明该项目是可行的。该应用程序概述了一个项目，以解决细胞 - 细胞粘附的非常困难但至关重要的方面 - 脱糖体中的蛋白质 - 蛋白质粘合剂相互作用。脱糖体是表皮中的主要粘附结构，它们将角质形成细胞粘合在一起。尽管它们的意义，但尚不清楚脱骨体如何形成表皮细胞之间的紧密，有序和动态的粘合剂联系。理解这个问题对于阐明严重的人类起泡皮肤疾病的机制，包括Pemphigus foliceus和Pemphigus vulgaris的机制。此外，这些知识将指定靶向降低表皮细胞之间凝聚力的化合物的靶标。这些化合物可能在伤口愈合中应用，并在整个皮肤中促进其他药物或疫苗的快速输送。由于脱粒体参与细胞内信号传导，因此对其粘合剂相互作用的操纵可能会带来调节细胞内信号的新途径，以纠正过度增殖，异常分化和其他皮肤异常。