Structural and Functional Analysis of Signaling Proteins in Dendritic Spines

树突棘信号蛋白的结构和功能分析

基本信息

批准号：
8118577
负责人：
Wolfgang Peti
金额：
$ 32.09万
依托单位：
BROWN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-30 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8118577
关键词：
AMPA Receptors Actins Binding Biochemical Biological Assay Brain Cells Chimeric Proteins Collaborations Complex Data Dendritic Spines Employee Strikes Event Glutamate Receptor Glutamates Goals Human body In Vitro In Vivo NMR Spectroscopy Individual Laboratories Mediating Medical Membrane Memory Modeling Molecular Morphology Movement Muscle N-Methyl-D-Aspartate Receptors N-Methylaspartate NMR Spectroscopy Neurabin Neurons Peptide Library Play Process Property Protein Binding Protein Serine/Threonine Phosphatase Protein phosphatase Protein-Serine-Threonine Kinases Proteins Regulation Resolution Role Scaffolding Protein Signal Transduction Signaling Protein Synapses Techniques Technology Tertiary Protein Structure Testing Universities base brain tissue density in vivo insight long term memory profilin protein protein interaction receptor scaffold spinophilin structural biology three dimensional structure trafficking transmission process

项目摘要

DESCRIPTION (provided by applicant): Signaling cascades direct information transmission and, in turn, function in processes as diverse as memory and muscle movement. These cascades are mediated by a network of highly specific and tightly regulated protein:protein interactions, including those made by serine/threonine kinases and serine/threonine phosphatases. Our long-term goal is to achieve an in-depth understanding of signaling networks with a special focus on serine/threonine phosphatase signaling, by using biochemical, biophysical, structural biology, especially NMR spectroscopy and in vivo technologies. In the proposed studies, we will focus on the role of the serine/threonine phosphatase Protein Phosphatase 1 (PP1) in dendritic spine signaling, as it is one of the most important protein phosphatases in brain tissue. Two of the major interaction partners of PP1 in neurons are the large, multi-domain scaffolding proteins spinophilin and neurabin. These proteins target PP1 to its cellular point of action, the post synaptic density of dendritic spines. This targeting of PP1 by spinophilin and neurabin is responsible for the PP1-mediated regulation of glutamatergic AMPA/NMDA receptor activity and trafficking. To understand the spinophilin:PP1 and neurabin:PP1 signaling networks in molecular detail, we will use NMR spectroscopy to elucidate the 3-dimensional structures and scaffolding properties of the spinophilin and neurabin protein interaction domains, both as isolated domains and in complex with their protein binding partners. Based on the results obtained from the NMR studies, we will investigate the in vivo functional significance of these interactions in cells and neurons. These in vivo studies will be done in close collaboration with the Nairn laboratory at Yale University. Furthermore, neurabin and spinophilin are capable of inducing cytoskeletal remodeling by means of modulating f-actin organization and by activity-induced profilin targeting. Activity-induced profilin targeting plays a major role in the stabilization of the dendritic spine morphology and thus long-term memory. We will first generate biochemical and structural in vitro data of these biologically critical events, and subsequently test our models using in vivo functional assays to provide a detailed understanding of critical parameters in dendritic spine signaling and morphology. These structural and functional studies will provide a detailed understanding of the roles of spinophilin and neurabin in brain and will, as a long term goal, enable us to selectively modulate these particular signaling cascades for medical benefit.

描述（由申请人提供）：信号级联直接信息传输，进而在记忆和肌肉运动等多样化的过程中发挥作用。这些级联反应是由高度特异性和紧密调控蛋白质的网络介导的：蛋白质相互作用，包括丝氨酸/苏氨酸激酶和丝氨酸/丝氨酸磷酸酶制成的蛋白质相互作用。我们的长期目标是通过使用生化，生物物理，结构生物学，尤其是NMR光谱和体内技术，以特别关注丝氨酸/苏氨酸磷酸酶信号传导的信号网络深入了解。在拟议的研究中，我们将重点关注丝氨酸/苏氨酸磷酸酶蛋白磷酸酶1（PP1）在树突状脊柱信号传导中的作用，因为它是脑组织中最重要的蛋白质磷酸酶之一。 PP1在神经元中的两个主要相互作用伙伴是大型多域脚手架蛋白自旋蛋白和神经蛋白酶。这些蛋白质靶向其细胞作用点，即树突状刺的突触密度。 Spinophilin和Neurabin对PP1的靶向是PP1介导的谷氨酸能AMPA/NMDA受体活性和运输的调节。为了理解分子细节的自旋粒蛋白：PP1和神经蛋白：PP1信号网络，我们将使用NMR光谱法来阐明自旋蛋白和神经蛋白蛋白质相互作用的3维结构和脚手架特性，既可以作为孤立的结构域，又与其蛋白质结合的蛋白质构造蛋白相关。根据从NMR研究获得的结果，我们将研究细胞和神经元中这些相互作用的体内功能意义。这些体内研究将与耶鲁大学的NAIRN实验室密切合作。此外，神经阿拉伯蛋白和旋转蛋白能够通过调节F-肌动蛋白组织和活性诱导的profilin靶向来诱导细胞骨架重塑。活性诱导的profilin靶向靶向在树突状脊柱形态的稳定中起着重要作用，因此长期记忆。我们将首先生成这些生物学上关键事件的生化和结构性的体外数据，然后使用体内功能测定法测试我们的模型，以详细了解树突状脊柱信号传导和形态中关键参数。这些结构和功能研究将提供详细的理解自旋素和神经蛋白在大脑中的作用，并将作为一个长期目标，使我们能够选择性地调节这些特定的信号级联以获得医疗益处。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Flexibility in the PP1:spinophilin holoenzyme.

PP1：亲旋蛋白全酶的灵活性。

DOI：
10.1016/j.febslet.2010.11.022
发表时间：
2011
期刊：
FEBS letters
影响因子：
3.5
作者：
Ragusa,MichaelJ;Allaire,Marc;Nairn,AngusC;Page,Rebecca;Peti,Wolfgang
通讯作者：
Peti,Wolfgang

Structural signature of the MYPT1-PP1 interaction.

MYPT1-PP1 相互作用的结构特征。

DOI：
10.1021/ja107810r
发表时间：
2011
期刊：
Journal of the American Chemical Society
影响因子：
15
作者：
Pinheiro,AndersonS;Marsh,JosephA;Forman-Kay,JulieD;Peti,Wolfgang
通讯作者：
Peti,Wolfgang

NIPP1 maintains EZH2 phosphorylation and promoter occupancy at proliferation-related target genes.

DOI：
10.1093/nar/gks1255
发表时间：
2013-01
期刊：
Nucleic acids research
影响因子：
14.9
作者：
Minnebo N;Görnemann J;O'Connell N;Van Dessel N;Derua R;Vermunt MW;Page R;Beullens M;Peti W;Van Eynde A;Bollen M
通讯作者：
Bollen M

Structure-function analysis of the filamentous actin binding domain of the neuronal scaffolding protein spinophilin.

神经元支架蛋白亲旋蛋白丝状肌动蛋白结合域的结构功能分析。