Proteomics of the Proteasome Interacting Networks

蛋白酶体相互作用网络的蛋白质组学

基本信息

批准号：
10427266
负责人：
Lan Huang
金额：
$ 35.33万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-08-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10427266
关键词：
26S proteasome Address Architecture Biochemical Biological Cells Chemistry Complex Data Detection Deterioration Development Disease Emerging Technologies Environment Excision Functional disorder Funding Future Goals Human Huntington Disease Huntington gene Huntington protein Hydrophobic Interactions Hydrophobicity Impairment In Vitro Investigation Link Mass Spectrum Analysis Mediating Methodology Molecular Molecular Conformation Monitor Nerve Degeneration Neurodegenerative Disorders Onset of illness Organism Peptides Pharmacology Phosphorylation Photochemistry Physiological Proteasome Inhibition Protein Conformation Protein Dynamics Protein-Protein Interaction Map Proteins Proteomics Role Structure Structure-Activity Relationship Sulfoxide System Technology Testing Therapeutic Time Work base crosslink density design effective therapy improved in vivo innovation insight misfolded protein multicatalytic endopeptidase complex mutant neuropathology novel novel strategies novel therapeutic intervention polyglutamine protein aggregation protein complex protein misfolding protein protein interaction restoration structural biology success

项目摘要

Huntington’s disease (HD) is one of the most devastating neurodegenerative disorders (NDs) that currently lacks effective therapies. Caused by toxic aggregation of mutant Huntingtin (Htt) proteins carrying abnormally long polyglutamine (polyQ) repeats, their timely removal through proteasomal degradation is critical for delaying onset of the disease. Given its crucial role as the central machine responsible for degradation of damaged and misfolded proteins such as Htt and other aggregation-prone proteins, 26S proteasome impairment has been recognized as one of the hallmarks of NDs associated with neuropathology. While it has been suggested that protein aggregates can induce conformational changes in the 26S to reduce its function, activation of proteasomes through phosphorylation appears to enhance the removal of Htt mutants. However, the molecular details underlying proteasome inhibition and activation in HD remain unclear. To address these unknowns, it is essential to quantitatively assess Htt aggregation and phosphorylation-dependent conformations of the 26S in cells to obtain a mechanistic understanding of the structure-function relationship of HD-associated proteasomes. Such investigations have remained previously unexplored due to lack of proper strategies. During the current funding cycle, we have demonstrated that cross-linking mass spectrometry (XL- MS) is effective for studying in vivo structural dynamics of proteasome complexes. While the development of specific residue-targeting MS-cleavable cross-linkers has further improved our capability to map protein-protein interactions (PPIs), interactions at hydrophobic regions remain difficult to characterize due to lack of targetable residues. Therefore, it is necessary to explore alternative chemistries for capturing structural details in those regions in order to comprehensively dissect proteasome conformational dynamics in cells. Here, we aim to develop photochemistry-based XL-MS platforms to enable their application for complex PPI mapping in vivo and in vitro. In addition, we intend to develop integrated QXL-MS platforms to define the temporal dynamics of the 26S proteasome upon Htt aggregation and phosphorylation, yielding molecular details to delineate the structure-function relationship of HD-impaired proteasomes. This project not only represents a great leap in XL-MS technology, but also helps address important yet unresolved biological questions associated with HD that have great potential for future therapeutic exploitation.

亨廷顿氏病（HD）是目前最具破坏性的神经退行性疾病（NDS）之一缺乏有效的疗法。由携带异常携带的突变体亨廷汀（HTT）蛋白的有毒聚集长聚谷氨酰胺（polyq）重复，它们及时通过蛋白酶体降解对延迟疾病发作。鉴于其作为负责退化的中央机器的关键作用受损和折叠的蛋白质，例如HTT和其他容易发生蛋白，26S蛋白酶体障碍被认为是与神经病理学相关的NDS的标志之一。虽然有建议蛋白质聚集体可以诱导26S中的构象变化以降低其功能，通过磷酸化激活蛋白酶体似乎增强了HTT突变体的去除。然而， HD中蛋白酶体抑制和激活的分子细节尚不清楚。解决这些未知数，必须定量评估HTT聚集和磷酸化依赖性细胞中26S的构象以获得对结构功能关系的机械理解 HD相关蛋白酶体。由于缺乏适当的适当策略。在当前的融资周期中，我们已经证明了交联的质谱法（XL- MS）对于研究蛋白酶体复合物的体内结构动力学有效。而发展特定居住的MS可裂解交联链链接进一步提高了我们映射蛋白质蛋白的能力相互作用（PPI），由于缺乏目标，疏水区域的相互作用仍然难以表征残留物。因此，有必要探索替代化学物质以捕获其中的结构细节为了全面剖析细胞中蛋白酶体会议动力学。在这里，我们的目标是开发基于光化学的XL-MS平台，以启用其用于体内复杂PPI映射的应用并在体外。此外，我们打算开发集成的QXL-MS平台，以定义 HTT聚集和磷酸化时的26S蛋白酶体，产生分子细节以描绘 HD损伤蛋白酶体的结构功能关系。这个项目不仅代表了一个巨大的飞跃 XL-MS技术，但也有助于解决与高清相关的重要但未解决的生物学问题这具有未来治疗剥削的巨大潜力。

项目成果

期刊论文数量（64）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Exploring an Alternative Cysteine-Reactive Chemistry to Enable Proteome-Wide PPI Analysis by Cross-Linking Mass Spectrometry.

DOI：
10.1021/acs.analchem.2c04986
发表时间：
2023-01-31
期刊：
ANALYTICAL CHEMISTRY
影响因子：
7.4
作者：
Jiao, Fenglong;Salituro, Leah J.;Yu, Clinton;Gutierrez, Craig B.;Rychnovsky, Scott D.;Huang, Lan
通讯作者：
Huang, Lan

Inhibitors of cullin-RING E3 ubiquitin ligase 4 with antitumor potential.

DOI：
10.1073/pnas.2007328118
发表时间：
2021-02-23
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Wu K;Huynh KQ;Lu I;Moustakim M;Miao H;Yu C;Haeusgen MJ;Hopkins BD;Huang L;Zheng N;Sanchez R;DeVita RJ;Pan ZQ
通讯作者：
Pan ZQ

Defining dynamic protein interactions using SILAC-based quantitative mass spectrometry.

使用基于 SILAC 的定量质谱法定义动态蛋白质相互作用。