Structural stabilization of Parkinson's disease linked mutant DJ-1

帕金森病相关突变体 DJ-1 的结构稳定

• 批准号: 8222821
• 负责人: Soumya Ray
• 金额: $ 26.78万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8222821
• 关键词: 1p36.2; Affect; Affinity; American; Binding; Biological Assay; Calorimetry; Cells; Cessation of life; Chemicals; Collaborations; Defect; Development; Dissociation; Disulfides; Dyes; Exercise; FDA approved; Genes; Goals; Human Genome; In Vitro; Investigation; Laboratories; Lead; Libraries; Ligand Binding; Ligands; Link; Location; Measurement; Mediating; Methods; Mitochondria; Modeling; Molecular Chaperones; Mutation; Nature; Neurodegenerative Disorders; Neurons; Neurosciences; Newly Diagnosed; Oxidative Stress; PARK7 gene; Parkinson Disease; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Property; Proteins; Reverse Transcriptase Polymerase Chain Reaction; Role; Scanning; Screening procedure; Signal Transduction; Structure; Structure-Activity Relationship; Testing; Therapeutic; Time; Titrations; Toxic effect; Work; alpha synuclein; analog; apoptosis inducing factor; base; design; disulfide bond; drug discovery; early onset; improved; loss of function mutation; melting; mutant; neuroprotection; novel; overexpression; oxidation; patient population; programs; public health relevance; small molecule; small molecule libraries; synuclein; synucleinopathy

DESCRIPTION (provided by applicant): Loss-of-function mutations such as A104T and M26I in the DJ-1 gene lead to structural destabilization and have been linked to early onset Parkinson's disease. We have recently demonstrated that structural defects in DJ-1 can be corrected by introduction of disulfide bridges that restore WT-like properties in the mutant protein. A pilot screen carried out using differential scanning fluorimetry using a library of 1080 FDA approved compounds yielded a number of molecules that improve the stability of mutant DJ-1. In addition, the same molecules increased the stability of WT DJ-1 as well. Wild type DJ-1 blocks a-synuclein aggregation, a property not shared by the familial mutants. However, the molecules discovered from the screen seem to restore chaperone like function in mutant DJ-1, as well as increase the ability of WT DJ-1 to function as a chaperone for extended periods of time. A structure activity relationship exercise carried out on one of the hits led to the identification of an analog that bound to DJ-1 with a dissociation constant of 480nM. Accurate dissociation constants were determined using isothermal titration calorimetry. The major goal of this proposal is to extend the screening to a larger chemical library to identify diverse classes of molecules for DJ-1, which can work as co-chaperones and block aggregation of a-synuclein. High affinity ligands will represent a starting for development of PD therapeutics, as well as probes for understanding the role of DJ-1 in cell based models in greater details. PUBLIC HEALTH RELEVANCE: Parkinson's disease (PD) is the second most common neurodegenerative disorder. It affects over 1 million Americans and more than 60,000 patients are newly diagnosed each year. Mutation in a recently identified gene product of unknown function, DJ-1, has been implicated in autosomal- recessive early onset PD. The mutations appear to compromise the chaperone like function of DJ-1. The major objective of this proposal is to identify small molecules that would restore normal function in mutants of DJ-1. In addition, these molecules would enhance the function of wild-type DJ-1 and, thereby, have an indirect effect on a greater population of patients with PD and other synucleinopathies.

描述（由申请人提供）：DJ-1基因中的功能丧失突变（如A104 T和M26 I）导致结构不稳定，并与早发性帕金森病有关。我们最近证明，DJ-1的结构缺陷可以通过引入二硫键来纠正，从而恢复突变蛋白中的WT样特性。使用1080种FDA批准的化合物的文库，使用差示扫描荧光法进行中试筛选，产生了许多改善突变体DJ-1稳定性的分子。此外，相同的分子也增加了WT DJ-1的稳定性。野生型DJ-1阻断α-突触核蛋白聚集，这是家族性突变体不具有的特性。然而，从筛选中发现的分子似乎恢复了突变体DJ-1中的分子伴侣样功能，以及增加WT DJ-1作为分子伴侣长时间发挥作用的能力。对其中一个命中进行的结构活性关系练习导致鉴定出与DJ-1结合的类似物，其解离常数为480 nM。准确的解离常数测定使用等温滴定量热法。该提案的主要目标是将筛选扩展到更大的化学文库，以鉴定DJ-1的不同类型的分子，其可以作为共分子伴侣并阻断α-突触核蛋白的聚集。高亲和力配体将代表开发PD治疗剂的起点，以及用于更详细地理解DJ-1在基于细胞的模型中的作用的探针。 公共卫生相关性：帕金森病（PD）是第二大最常见的神经退行性疾病。它影响了超过100万美国人，每年新诊断出超过60，000名患者。最近鉴定的功能未知的基因产物DJ-1中的突变与常染色体隐性早发性PD有关。这些突变似乎损害了DJ-1的伴侣样功能。该提案的主要目标是鉴定能够在DJ-1突变体中恢复正常功能的小分子。此外，这些分子将增强野生型DJ-1的功能，从而对更多的PD和其他突触核蛋白病患者群体产生间接影响。