Arrestin interactions with non-receptor binding partners

抑制蛋白与非受体结合伙伴的相互作用

基本信息

批准号：
8458058
负责人：
VSEVOLOD V. GUREVICH
金额：
$ 28.51万
依托单位：
VANDERBILT 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：
2007-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8458058
关键词：
Affect Affinity Alzheimer&apos s Disease Apoptosis Apoptotic Arrestins Binding Biochemical Biological Assay Cell Death Cell Proliferation Cell Survival Cells Cessation of life Co-Immunoprecipitations Complex Data Disease Dominant-Negative Mutation Drug Targeting Equilibrium Exclusion Family G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors GTP-Binding Proteins Human Individual Label Life Link MAP Kinase Gene MAP Kinase Modules MAPK10 gene MAPK8 gene Malignant Neoplasms Mediating Methods Mitogen-Activated Protein Kinases Molecular Mutagenesis Neurodegenerative Disorders Nuclear Parkinson Disease Pathway interactions Peptides Phosphotransferases Play Probability Research Role Signal Pathway Signal Transduction Signaling Protein Site Spectrum Analysis Testing Therapeutic Time X-Ray Crystallography arrestin3 base design interest mutant non-visual arrestins novel protein protein interaction receptor receptor binding receptor internalization response scaffold small molecule tool tumorigenesis upstream kinase

项目摘要

DESCRIPTION (provided by applicant): Arrestins were first described as negative regulators of G protein-coupled receptor (GPCR) signaling via G proteins. New data show that the free and receptor-bound arrestins initiate signaling through MAP kinases, which regulate cell death, survival, and proliferation. In particular, both free and receptor- associated arrestin-3 scaffolds ASK1-MKK-JNK cascade, promoting JNK activation. Here we propose to elucidate the structural basis of arrestin-dependent activation of pro-apoptotic JNK family kinases and their activators MKK4/7 using biochemical and biophysical methods. The molecular mechanisms of the assembly of multi-protein signaling complexes (signalosomes) organized by arrestin-3 will be established, and arrestin-3 residues critical for JNK activation will be identified. Based on ths info, arrestin-3 mutants that bind ASK1, MKK and JNK, but do not promote JNK activation will be constructed. The potential of arrestin mutants with dramatically reduced ability to activate JNKs, several of which we already have, to protect cells against insults and prolong their survival will be tested. We have also constructed arrestin-3 mutants that activate JNKs more efficiently than parental wild type arrestin-3. We established the paradigm where arrestin-3-dependent JNK activation plays key role in cell survival. We will test the ability of hyperactive arrestin-3 mutants to facilitate cell death. We showed that inactive mutants tie up JNKs and upstream kinases in unproductive complexes, thereby acting in dominant- negative manner. We will test the potential of these mutants to protect cells and prolong their survival. Molecular toos that specifically increase or block pro-apoptotic signaling have therapeutic potential in disorders associated with excessive cell proliferation (e.g., cancer) or death (e.g., neurodegenerative diseases).

描述（由申请人提供）：首先将阻止蛋白描述为G蛋白偶联受体（GPCR）信号的负调节剂，通过G蛋白传导。新的数据表明，自由和受体结合的停滞蛋白通过MAP激酶启动信号传导，该MAP激酶调节细胞死亡，生存和增殖。特别是，免费和与受体相关的逮捕素-3脚手架 ask1-mkk-jnk级联，促进JNK激活。在这里，我们建议使用生物化学和生物物理方法阐明促凋亡JNK家族激酶及其激活剂MKK4/7的结构基础。将确定由抑制蛋白3组织的多蛋白信号传导复合物（信号体）组装的分子机制，将确定对JNK激活至关重要的抑制蛋白3残基。基于THS信息，将构建结合Ask1，MKK和JNK但不会促进JNK激活的逮捕蛋白-3突变体。具有大幅度降低激活JNK的能力的抑制蛋白突变体的潜力，我们已经拥有了几种，可以保护细胞免受损害并延长其存活率。我们还构建了逮捕蛋白3突变体，该突变体比父母野生型逮捕更有效地激活JNK。我们建立了范式，其中抑制素3依赖性JNK激活在细胞存活中起关键作用。我们将测试多动抑制蛋白3突变体促进细胞死亡的能力。我们表明，非活性突变体将JNK和上游激酶在非生产性的复合物中绑定，从而以显性负面方式作用。我们将测试这些突变体保护细胞并延长其存活的潜力。特异性增加或阻断促凋亡信号传导具有治疗潜力的分子Toos 与过度细胞增殖（例如癌症）或死亡（例如神经退行性疾病）有关。