Adhesive & Morphogenetic Role of a Xenopus Protocadherin

粘合剂

• 批准号: 6619702
• 负责人: XUEJUN CHEN
• 金额: $ 4.64万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6619702
• 关键词: SDS polyacrylamide gel electrophoresis; Xenopus; affinity chromatography; biological signal transduction; cadherins; calcium ion; cell adhesion; cell adhesion molecules; cell motility; cellular polarity; cytoplasm; dimer; embryogenesis; gel filtration chromatography; histogenesis; immunofluorescence technique; polymerase chain reaction; postdoctoral investigator; protein protein interaction; tissue /cell culture; transfection /expression vector; western blottings; yeast two hybrid system

DESCRIPTION (provided by applicant): Protocadherins are new members of the cadherin superfamily. Despite the great number of protocadherin genes identified in vertebrates, little is known about their biochemical properties, functions or working mechanisms. Xenopus paraxial protocadcadherin (PAPC) was recently shown to play a role in morphogenetic cell movements during early embryonic development. However, the adhesive properties of PAPC still remain ambiguous and poorly understood, and whether its morphogenetic role is realized through its adhesive function or by other mechanisms is unclear. The long-term goals of this proposal are to elucidate the adhesive functions and mechanisms of PAPC and to understand the mechanisms of its morphogenetic activity by identifying the cytoplasmic PAPC-binding proteins. First, the adhesive properties and mechanisms, including adhesive strength, specificity, lateral dimerization and clustering, will be examined and analyzed with the cell aggregation, bead aggregation and cell attachment assays; second, the cellular localization of PAPC will be monitored to assess its role in polarized cell movements; third, the function of PAPC cytoplasmic domain, especially a highly conserved region, on the adhesive and morphogenetic activity of PAPC will be analyzed using deletion mutants; and finally, the proteins that bind PAPC cytoplasmic tail will be identified as candidates for regulation of PAPC adhesive function or PAPC-mediated signaling by means of affinity chromatography and/or the yeast two-hybrid method. The proposed research will shed light on the role of protocadherins in embryonic development and should provide important suggestions to the treatment of birth defects and other developmental diseases.

描述（由申请人提供）：原钙粘蛋白是钙粘蛋白超家族的新成员。尽管有大量的原钙粘蛋白基因 在脊椎动物中发现，但对其生化特性知之甚少， 职能或工作机制。非洲爪蟾近轴原钙粘蛋白 (PAPC) 最近显示在早期形态发生细胞运动中发挥作用 胚胎发育。然而，PAPC 的粘合性能仍然存在 模棱两可且知之甚少，以及它的形态发生作用是否实现 通过其粘合功能或通过其他机制尚不清楚。长期来看 该提案的目标是阐明粘合功能和机制 PAPC 并了解其形态发生活性的机制 鉴定细胞质 PAPC 结合蛋白。一、粘合剂 性质和机制，包括粘合强度、特异性、横向 二聚化和聚类，将用细胞进行检查和分析 聚集、珠聚集和细胞附着测定；第二，蜂窝 将监测 PAPC 的定位以评估其在极化细胞中的作用 动作；第三，PAPC胞质结构域的功能，特别是高度 保守区域，对 PAPC 的粘附和形态发生活性的影响将是 使用缺失突变体进行分析；最后，结合 PAPC 的蛋白质 细胞质尾部将被确定为 PAPC 调控的候选者 粘附功能或 PAPC 介导的亲和力信号传导 色谱法和/或酵母二杂交法。拟议的研究将 阐明原钙粘蛋白在胚胎发育中的作用，应该 为出生缺陷等的治疗提供重要建议 发育性疾病。