Structure And Mechanism In Intracellular Notch Signaling

描述：Noch信号转导是一种普遍的跨膜信号转导。 对细胞命运的决定、发育、 真核生物的动态平衡和神经系统功能。人类的遗传损伤 途径与人类疾病状态有关，如癌症和神经疾病 精神错乱。该途径的中心成分是胞浆-1000残基。 Notch跨膜受体；在激活该途径后，这 受体片段是由一种蛋白酶释放的，并增强下游 信号事件。这种称为NIC的可溶片段的活性是 由至少九种细胞内效应蛋白通过 被认为是直接的互动。因此，NIC是一个复杂的 互动。 在这里提出的研究的目的是阐明和量化 该网络如何调节的分子机制和结构细节 并决定细胞的命运。巴里克博士将确定和描述 通过溶液分析得到NIC沿线的结构和功能域 猪源嵌套性多肽和重叠多肽的结构与稳定性 网卡。还将确定细胞内的结构域图 效应器。然后这些域将被用来分析成对的和更高阶的 互动。系统的高吞吐量方法将进行定性筛选 对于所有可能的成对相互作用；传统溶液热力学 将使用各种方法来量化成对互动的强度。Dr。 巴里克将研究现有Notch途径突变对结构的影响 稳定性和成对相互作用，以帮助将体外结果与 信号通路的生物学。多蛋白质变构相互作用的研究进展 我们的NIC和效应域将被识别和量化；阐明 这种相互作用将有助于阐明调控的分子机制 凹槽信号。最后，巴里克博士将明确结构域和 复合体，通过以下方式提供结构域和复合体的原子级描述 X射线结晶学。

DESCRIPTION: Notch signaling is a general transmembrane signal transduction pathway that is essential for cell fate determination, development, homeostasis, and nervous system function in eukaryotes. Genetic lesions in the pathway are linked to human disease states such as cancer and neurological disorders. A central component of this pathway is the cytosolic -1000 residues of the Notch transmembrane receptor; upon activation of the pathway, this receptor fragment is released by a protease, and potentiates downstream signaling events. The activity of this soluble fragment, termed NIC, is modulated by at least nine intracellular effector proteins through what is thought to be direct interactions. Thus, NIC is the hub of a complex network of interactions. The aim of the research proposed here is to elucidate and quantify the molecular mechanisms and structural details of how this network modulates signaling and determines cell fate. Dr. Barrick will identify and characterize the structural and functional domains along NIC through analysis of solution structure and stability of nested and overlapping polypeptides derived from NIC. Structural domain maps will also be determined for intracellular effectors. These domains will then be used to analyze pairwise and higher order interactions. A systematic high-throughput approach will screen qualitatively for all possible pairwise interactions; conventional solution thermodynamic methods will be used to quantify the strength of pairwise interactions. Dr. Barrick will study the effect of existing Notch pathway mutations on structural stability and pairwise interactions, to help connect in vitro results to the biology of the signaling pathway. Multi-protein allosteric interactions among our NIC and effector domains will be identified and quantified; elucidation of such interactions will help to clarify the molecular mechanisms that modulate Notch signaling. Finally, Dr. Barrick will crystallize structural domains and complexes to provide an atomic level description of domains and complexes by x-ray crystallography.