Eph and Lyn hyper-phosphorylation and CRMP interactions in AD"

AD中Eph和Lyn过度磷酸化与CRMP相互作用"

基本信息

批准号：
10746170
负责人：
MATTHIAS BUCK
金额：
$ 24.15万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-15 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10746170
关键词：
Actins Affinity Alzheimer&apos s Disease Amyloid beta-Protein Antibodies Antigens Binding Biochemical Bioinformatics Biophysics Brain C-terminal Cardiovascular system Cells Complex Computer Models Cytoskeleton Data Development Diagnostic Disease Docking Early Diagnosis Eph Family Receptors Exploratory/Developmental Grant Family Family member Future Future Generations Grain Homologous Gene In Vitro Knowledge Learning Link Mammalian Cell Mediator Memory Microtubules Molecular Mutagenesis Mutation Natural regeneration Nature Neurodegenerative Disorders Neuronal Injury Neurons Organism Parkinson Disease Pattern Peptides Phosphorylation Phosphotransferases Preparation Process Protein Kinase Proteins Research Research Personnel Risk Role SAM Domain Sampling Semaphorin-3A Site Structure System Therapeutic Therapeutic Intervention Time Validation aging brain antibody diagnostic biophysical techniques cell motility design experimental study follow-up glycogen synthase kinase 3 beta insight mimetics molecular dynamics molecular modeling neuron development plexin potential biomarker protein complex protein purification reconstitution response scaffold structural biology targeted treatment tau Proteins tool

项目摘要

Eph receptors make guidance decisions for cell migration in cardiovascular and neuronal development, disease, and regeneration. Eph receptors are also involved with higher brain functions, memory and learning. A protein that has been associated with Alzheimer’s, Parkinson’s and other neuronal diseases and injuries is the Collapsin Response Mediator Protein (CRMP) which interacts with several kinases and becomes hyper-phosphorylated alongside increased activation of the kinases. The hyper- phosphorylated CRMP then disrupts the formation of actin and microtubule cytoskeletal structures and it thought to impede Aβ and tau clearance. Cytosolic Lyn kinase, a close homologue of Fyn kinase, is known to interact directly with EphA4 and here, for the first time, we show that an EphA family member and EphB2 directly interacts with CRMP. Analogous to another guidance and cell migration system, the Fyn-Plexin-CRMP complex, the Lyn-EphA4-CRMP association is likely to form a complex with Cdk5 and GSK3β kinases, each also involved in Alzheimer’s. The features of the Eph-CRMP and Eph-Lyn interacting interfaces need to be characterized, as they will be potential biomarkers and targets for therapeutic intervention. The proposal has two main aims. Aim 1) seeks to establish the in vitro phosphorylation patters of various kinases on the intracellular domains of EphA4 and –B2 in the presence and absence of CRMP and to further validate the formation of the complex in cells. Aim 2) The effect that the corresponding phosphomimetic/phospho-defective mutations have on the level of activity on these Eph receptor intracellular regions and of the kinases will be studied with purified proteins in vitro. Eventually, using the knowledge from this project, Antibodies or complex-disrupting-peptides may drive the future development of early detection diagnostics and/or therapeutics.

EPH受体在心血管和神经元发育中为细胞迁移做出指导决策，疾病和再生。 EPH受体还涉及更高的大脑功能，记忆力和学习。与阿尔茨海默氏症，帕金森氏症和其他神经元疾病有关的蛋白质损伤是与几种激酶相互作用的折叠蛋白反应介质蛋白（CRMP）并随着激酶激活的增加而变得高磷酸化。超级然后，磷酸化的CRMP破坏了肌动蛋白和微管细胞骨架结构的形成，IT 想想阻碍Aβ和TAU清除。 Fyn激酶的紧密同源物的胞质Lyn激酶是已知可以直接与Epha4互动，在这里，我们首次表明了Epha家族成员 EPHB2直接与CRMP相互作用。类似于另一个指导和细胞迁移系统， Fyn-Plexin-CRMP复合物，Lyn-Epha4-Crmp关联可能与CDK5形成复合物，并且 GSK3β激酶也参与阿尔茨海默氏症。 EPH-CRMP和EPH-LYN的特征相互作用的接口需要表征，因为它们将是潜在的生物标志物和目标治疗干预。该提案有两个主要目标。目标1）试图建立体外在Epha4和–b2的细胞内结构域中各种激酶的磷酸化模式 CRMP的存在和不存在，并进一步验证复合物在细胞中的形成。目标2）相应的磷酸/磷酸缺陷突变对活性水平的影响在这些EPH受体上，细胞内区域和激酶将在纯化的蛋白体外。最终，使用该项目的知识，抗体或复杂的干扰肽可能推动未来的早期检测诊断和/或治疗的发展。