Novel assay for ubiquitin pathway enzymes as biomarkers

泛素途径酶作为生物标志物的新检测方法

• 批准号: 7480799
• 负责人: Tauseef R. Butt
• 金额: $ 26.73万
• 依托单位: LIFESENSORS, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7480799
• 关键词: Alzheimer' s Disease; Basic Science; Binding; Biological Assay; Biological Markers; Biology; Cell physiology; Cellular Structures; Chromosome Pairing; Class; Cleaved cell; Complex; Deubiquitinating Enzyme; Development; Diagnosis; Diagnostic; Disease; Environment; Enzymes; Excision; Family; Frontotemporal Lobar Degenerations; Functional disorder; Generations; Goals; Hand; Human Genome; Lead; Libraries; Light; Link; Lysine; Malignant Neoplasms; Mediator of activation protein; Medical; Monitor; Nervous System Physiology; Neurodegenerative Disorders; Neurons; Pathogenesis; Pathology; Pathway interactions; Peptide Library; Personal Satisfaction; Pharmaceutical Preparations; Phase; Play; Protein Family; Protein Subunits; Proteins; Proteolysis; Public Health; Purpose; Reagent; Regulation; Reporter; Research Personnel; Role; Sclerosis; Solutions; Specificity; Surface; Synapses; Technology; Therapeutic Uses; Ubiquitin; Ubiquitin Like Proteins; Western Blotting; alpha synuclein; assay development; clinical application; commercial application; drug discovery; enzyme activity; enzyme pathway; enzyme substrate; inhibitor/antagonist; nervous system disorder; new technology; novel; preference; tau Proteins; tool; ubiquitin ligase

DESCRIPTION (provided by applicant): It has been well established that the ubiquitin pathway plays an important role as a key regulator of neuronal biology. The ubiquitin pathway enzymes are essential and critical mediators of synaptic adaptability in mature neurons, and dysfunctional ubiquitin pathway enzymes or regulation is associated with diverse neurodegenerative disorders. Deubiquitinating enzymes, or DUBs belong to a family of ~100 primarily single subunit proteins that remove ubiquitin from lysine residues of target proteins. Evidence suggests that that dysregulated or un-regulated DUB(s) can lead to neurodegenerative disease. Most commonly used assays for DUB activity are not suitable for either clinical applications or drug discovery. In this proposal, a novel assay for the DUB family of proteins will be developed. The assay involves a semi-synthetic approach to develop reagents that are extensions of the C-terminus of ubiquitin that is conjugated to target proteins. A library of peptides can be extended from the C-terminus of ubiquitin; the best reagent for a given DUB can be selected from the library since classes of DUBs show different specificity preferences. In contrast to their fate in solution phase assays, entities cleaved by DUBs will be immobilized on a surface, allowing them to be suitable reagents for immobilized DUBs in a multiplex environment. Since the assay can identify a protein as a DUB, it can also uncover DUB interactive proteins or the accumulation of ubquitinated substrates common in neurodegenerative diseases. Thus, the assay will be useful for diagnostic as well as drug discovery purposes. In Phase II, the assay will be expanded to include all known DUBs. The commercial application of this technology is the generation of products for diagnostic and therapeutic uses as applied to DUB-linked medical needs such as neurodegenerative diseases. Proteins modified by another small protein called ubiquitin are key regulators of neuronal biology. Enzymes that catalyse the addition or removal of ubiquitin to target proteins are important regulators of neuronal cell function and are potentially useful for diagnosing various neurological diseases or discovering drugs to treat the diseases. PUBLIC HEALTH RELEVANCE: LifeSensors proposes to develop an assay for one of the major types of ubiquitin pathway enzymes known as DUBs (deubiquitinating enzymes). This assay represents a novel technology that will have potential utility in diagnostics and in the discovery of potent and selective drugs to treat Alzheimer's Disease and other neurological diseases.

描述（由申请人提供）：已经很好地确定，泛素途径是神经元生物学的关键调节剂的重要作用。泛素途径酶是成熟神经元中突触适应性的必不可少的介体，功能障碍的泛素途径酶或调节与多种神经退行性疾病有关。去泛素化酶或配音属于一个〜100个家族，主要是单个亚基蛋白，这些蛋白从靶蛋白的赖氨酸残基中去除泛素。证据表明，失调或未受监管的DUB会导致神经退行性疾病。 DUB活动的最常用测定法不适合临床应用或药物发现。在此提案中，将开发针对蛋白质配音家族的新颖测定法。该测定法涉及一种半合成方法，以开发是泛素C-末端的试剂，该试剂与靶蛋白结合在一起。可以从泛素的C末端扩展肽库；可以从库中选择给定配音的最佳试剂，因为配音的类别显示出不同的特异性偏好。与在溶液相测定中的命运相反，将DUB裂片的实体固定在表面上，从而使其成为在多重环境中固定的DUB的合适试剂。由于该测定法可以识别蛋白质为配音，因此它也可以发现DUB的互动蛋白或在神经退行性疾病中常见的紫外线底物的积累。因此，该测定对于诊断和药物发现目的将很有用。在第二阶段，该测定法将扩展到包括所有已知的DUB。该技术的商业应用是用于诊断和治疗用途的产品，这些产品应用于DUB连接的医疗需求，例如神经退行性疾病。由另一种称为泛素的小蛋白质修饰的蛋白质是神经元生物学的关键调节剂。催化泛素在靶蛋白中催化或去除泛素的酶是神经元细胞功能的重要调节剂，并且可能有助于诊断各种神经疾病或发现治疗疾病的药物。公共卫生相关性：救生员建议为一种称为DUBS（去泛素化酶）的主要类型的泛素途径酶之一开发一种测定法。该测定法代表了一种新型技术，该技术将在诊断和发现有效和选择性药物方面具有潜在效用，以治疗阿尔茨海默氏病和其他神经系统疾病。