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

基本信息

批准号：
8111537
负责人：
Soumya Ray
金额：
$ 22.28万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-04-01 至 2013-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8111537
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 small molecule small molecule libraries synuclein

项目摘要

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，例如A104T和M26I导致结构性不稳定，并与早期发作帕金森氏病有关。我们最近证明，DJ-1中的结构缺陷可以通过引入恢复突变蛋白中WT样性质的二硫键来纠正。使用1080 FDA批准化合物的库进行荧光示意图进行的试验屏幕产生了许多分子，可改善突变体DJ-1的稳定性。另外，相同的分子也提高了WT DJ-1的稳定性。野生型DJ-1阻断了a-核蛋白聚集，这是家族性突变体没有共享的特性。但是，从屏幕上发现的分子似乎可以恢复突变体DJ-1中的伴侣功能，并提高了WT DJ-1在长时间内起伴侣的能力。在其中一次命中进行的结构活动关系运动导致识别与DJ-1的模拟识别，其解离常数为480nm。使用等温滴定量热法确定精确的解离常数。该提案的主要目的是将筛选扩展到较大的化学文库，以识别DJ-1分子的各种类别，这些分子可以用作共伴侣并阻止A-核蛋白的聚集。高亲和力配体将代表开发PD疗法的开始，以及在更多细节中了解DJ-1在基于细胞模型中的作用的探针。公共卫生相关性：帕金森氏病（PD）是第二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60,000多名患者。最近鉴定出的未知功能DJ-1基因产物中的突变与常染色体引起的早期发作PD有关。突变似乎损害了DJ-1的伴侣伴侣的功能。该建议的主要目的是确定将恢复DJ-1突变体正常功能的小分子。此外，这些分子将增强野生型DJ-1的功能，从而对PD和其他突触核病患者群体有间接影响。