Assembly, dynamics and evolution of cell-cell and cell-matrix adhesions

描述（由申请人提供）：细胞粘附复合物是一类重要的生物医学多蛋白组装体。它们参与感知细胞与其外部环境之间的相互作用，然后启动和调节控制细胞迁移、细胞形状和功能组织、增殖和存活以及基因表达的细胞内信号。它们在进化上是古老的，对正常发育和体内平衡至关重要，并且在遗传疾病和癌症中有缺陷。 目前，缺乏协调一致的努力，整合可用的基因组（进化）和结构信息，严格解决这些多蛋白质复合物在原子，介观和宏观尺度的层次结构组织。进展的另一个障碍是用于多蛋白质复合物重构的大量纯化蛋白质的可用性，因为经典方法由于复杂蛋白质组装体的不稳定性质而不可行。 该联盟将利用大量目标蛋白质家族和信号网络的高通量表达和结构，以了解细胞-细胞和细胞-ECM粘附复合物的结构和功能组织。该联盟将整合结构生物学（X射线晶体学、原位复合物的电子显微镜）、生物化学和细胞生物学（体外）方面的专门知识 重建），化学和活细胞成像（原位生物传感器和笼蛋白）： 具体目标1：定义溶液中的多蛋白质相互作用、化学计量和亲和力（Liddington，Weis）。具体目标2：在生物膜上重构多蛋白复合物（Ginsberg，纳尔逊）。具体目标3：在生理环境中原位确定多蛋白复合物的结构（Hanein，Rehmann）。具体目标4：分析活细胞中动态蛋白质-蛋白质相互作用和组装（Hahn）。 公共卫生相关性：这是人类健康的一个关键领域。细胞粘附复合物在正常细胞和组织功能的各个方面都很重要，并且在遗传疾病和癌症中通常是有缺陷的。这些结果将为缺陷如何破坏正常功能并导致疾病提供新的理解，并可能确定新的治疗靶点。

DESCRIPTION (provided by applicant): Cell adhesion complexes are a bio-medically important class of multi-protein assemblies. They are involved in sensing Interactions between cells and their external environment, and then initiating and regulating intracellular signals that control cell migration, cell shape and functional organization, proliferation and survival, and gene expression. They are evolutionarily old, critical for normal development and homeostasis, and are defective in genetic diseases and cancer. Currently, there is a lack of a concerted effort to integrate available genomic (evolutionary) and structural information to rigorously solve the hierarchical structural organization of these multi-protein complexes at the atomic, meso and macro scale. A further barrier to progress is the availability of large amounts of purified proteins for multi-protein complex reconstitution since classical approaches are not feasible due to the unstable nature of complex protein assemblies. The Consortium will leverage high-throughput expression and structures of large sets of target families of proteins and signaling networks to understand the structural and functional organization of both cell-cell and cell-ECM adhesion complexes. The Consortium will integrate expertise in structural biology (X-ray crystallography, electron microscopy of in situ complexes), biochemistry and cell biology (in vitro reconstitution), chemistry and live cell imaging (in situ bio-sensors and caged proteins) by: Specific Aim 1: Define multi-protein interactions, stoichiometries and affinities in solution (Liddington, Weis). Specific Aim 2: Reconstitute multi-protein complexes on biological membranes (Ginsberg, Nelson). Specific Aim 3: Define structures of multi-protein complexes in situ, at a physiological environment (Hanein, Volkmann). Specific Aim 4: Analyze dynamic protein-protein interactions and assembly in live cells (Hahn). PUBLIC HEALTH RELEVANCE: This is a critical area of Human Health. Cell adhesion complexes are important in all aspects of normal cell and tissue function, and are commonly defective in genetic diseases and cancers. Results will provide new understanding of how defects disrupt normal function and contribute to disease, and potentially identify new therapeutic targets.