Development of chain selective polyubiquitin markers as early detectors of Parkinson’s and Alzheimer’s Disease

开发链选择性多聚泛素标记作为帕金森病和阿尔茨海默病的早期检测器

基本信息

批准号：
9406658
负责人：
Tauseef R. Butt
金额：
$ 19.03万
依托单位：
LIFESENSORS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9406658
关键词：
Affect Affinity Alzheimer&apos s Disease Applications Grants Architecture Autoimmune Diseases Autophagocytosis Binding Biochemical Biology C-terminal Cancer Center Cell Line Cells Chemicals Communicable Diseases Communities Complication Custom DNA Repair Detection Development Disease Disease model Doctor of Medicine Down-Regulation Drug Targeting Early Diagnosis Endocytosis Engineering Enzymes Excision Exhibits Functional disorder Glycine Goals Grant Half-Life Histones Immune response Inflammation Link Lysine Malignant Neoplasms Mammalian Cell Maps Mediating Modification Mono-S N-terminal Names Nature Neurodegenerative Disorders Neuronal Dysfunction Neurons Outcome Oxidative Stress Parkinson Disease Pathogenesis Pathway interactions Pattern Phase Phosphorylation Play Polyubiquitin Post-Translational Protein Processing Protein Microchips Proteins Proteomics Reagent Research Role Signal Transduction Specificity Staining method Stains Testing Texas Therapeutic Time Tube Ubiquitin Ubiquitination Universities Western Blotting amino group biological adaptation to stress biophysical techniques carboxylate design detector drug discovery experience experimental study improved misfolded protein multicatalytic endopeptidase complex neuron development neuron loss novel professor protein degradation protein function protein structure protein transport receptor receptor recycling response screening success tool ubiquitin mediated proteasome degradation

项目摘要

Mammalian cells remove misfolded proteins by degradation mediated by the ubiquitin (Ub) proteasome pathway and autophagy. The first step in the development of neuronal dysfunction or neuronal death is the lack of a response to misfolded proteins. Dysfunction in the Ub proteasome pathway in neurons leads to the accumulation of aggregates and plaques in neurons. Ubiquitylation of proteins not only controls the half-lives of proteins but also acts as a regulatory modification, e.g., histone ubiquitylation. Due to the dynamic nature of Ub conjugation and deconjugation, ubiquitylated proteins are extremely difficult to isolate and study. Identifying a protein with one particular Ub post translational modification out of the pool of modified proteins becomes nearly impossible without selective tools. A further complication is the presence of multiple types of Ub-Ub linkages in polyubiquitin chains. Ubiquitin is attached, via isopeptide bonds, to lysine residues in the target protein. These Ub-moieties can then serve as substrates for the conjugation of additional Ubs, again through the formation of isopeptide bonds between the C-terminus of one Ub and any of seven (7) lysines, or N-terminus of the target Ub. Ub chains with different linkages convey different messages to cells and, hence, determine the ultimate fate of the protein -- degradation, translocation, and oxidative stress response, to name a few. The precise information encoded in the various chain linkages is largely unknown due to a lack of reagents that selectively recognize these linkages. The goal of this proposal is to develop tools that allow the selective identification, quantification, and isolation of proteins modified by mono and polyubiquitin chains containing difficult to study linkages. This will be accomplished using a novel Ub binding microarray that contains all of the known Ub binding motifs from various proteins. In Phase I, we will identify and characterize novel Ub binding domains (UbDs) exhibiting selectivity for monoubiquitin, monoubiquitylated substrates and specific polyubiquitin linkages. Given the fact that dysregulation of the Ub proteasome pathway and autophagy have been strongly implicated as a first step in neurodegenerative diseases, the development of sensitive and novel chain selective tools will lead to early diagnosis of Alzheimer’s and Parkinson’s diseases (phase II).

哺乳动物细胞通过泛素（UB）蛋白酶体途径介导的降解去除错误折叠的蛋白质和自噬。神经元功能障碍或神经元死亡发展的第一步是缺乏对错误折叠蛋白的反应。 UB蛋白酶体途径的功能障碍神经元导致积累神经元中的聚集体和斑块的粒子。蛋白质的泛素化不仅控制蛋白质的半衰期，而且控制还充当调节性修饰，例如组蛋白泛素化。由于UB共轭的动态性质和脱糖，泛素化蛋白很难分离和研究。鉴定蛋白质一个特定的UB后翻译后的修改几乎是不可能的没有选择性工具。另一个并发症是多泛素中存在多种类型的UB-UB链接链。泛素通过异肽键连接到赖氨酸保留在靶蛋白中。这些UB-Moieties 然后，可以通过形成异肽来充当额外ubs缀合的基材一个UB的C端与七（7）个赖氨酸中的任何一个或目标Ub的N端之间的键之间的键。 UB链随着不同的链接传达给细胞的不同信息，因此确定蛋白质的最终命运 - 降解，易位和氧化应激反应仅举几例。编码的精确信息在各种链条中，由于缺乏选择性识别这些试剂，因此在很大程度上是未知的链接。该建议的目的是开发允许选择性识别，量化和通过很难研究联系的单声道和多泛素链修饰的蛋白质的分离。这将是使用新型UB结合微阵列完成蛋白质。在第一阶段，我们将识别并表征新型的UB结合域（UBD），表现出选择性单喹素，单纤维素，单喹素系统和特定的多泛素链接。鉴于事实 UB蛋白酶体途径和自噬的失调已被强烈牵连为第一步神经退行性疾病，敏感和新型链选择工具的开发将导致早期诊断阿尔茨海默氏症和帕金森氏病（第二阶段）。