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 A104T 和 M26I 等功能丧失突变会导致结构不稳定，并与早发帕金森病有关。我们最近证明，DJ-1 中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类似 WT 的特性。使用差示扫描荧光法使用 1080 种 FDA 批准的化合物库进行的初步筛选产生了许多提高突变体 DJ-1 稳定性的分子。此外，相同的分子还提高了 WT DJ-1 的稳定性。野生型 DJ-1 阻断 a-突触核蛋白聚集，这是家族突变体所不具备的特性。然而，从筛选中发现的分子似乎恢复了突变体 DJ-1 中的伴侣样功能，并增加了 WT DJ-1 长时间充当伴侣的能力。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 480nM。使用等温滴定量热法测定准确的解离常数。该提案的主要目标是将筛选扩展到更大的化学库，以识别 DJ-1 的不同类别的分子，这些分子可以作为共伴侣并阻止 α-突触核蛋白的聚集。高亲和力配体将代表开发 PD 疗法的起点，以及更详细地了解 DJ-1 在细胞模型中的作用的探针。公众健康相关性：帕金森病 (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，每年新诊断出超过 60,000 名患者。最近发现的功能未知的基因产物 DJ-1 的突变与常染色体隐性早发性帕金森病有关。这些突变似乎损害了 DJ-1 的伴侣蛋白功能。该提案的主要目标是鉴定能够恢复 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，从而对更多的 PD 和其他突触核蛋白病患者产生间接影响。