Safeguards of the Proteome: Elucidating the Roles of Protein Disaggregases

蛋白质组的保障：阐明蛋白质解聚酶的作用

• 批准号: 10460941
• 负责人: Meredith E. Jackrel
• 金额: $ 39.38万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10460941
• 关键词: Amyloid; Cardiovascular Diseases; Cells; Cellular Stress; Disease; Ensure; Goals; Health; Human; Life; Mass Spectrum Analysis; Neurodegenerative Disorders; Play; Protein Conformation; Proteins; Proteome; Research; Role; Stress; Substrate Specificity; System; Techniques; Technology; Testing; Therapeutic; Work; Yeasts; conformer; disease phenotype; human disease; insight; misfolded protein; new technology; novel therapeutics; prevent; programs; protein folding; protein misfolding; proteostasis; restoration

Abstract: The proper folding of proteins is essential for all life, making aberrant protein folding severely problematic. The protein homeostasis (proteostasis) network is a tightly regulated system that ensures that proteins are folded and degraded as necessary. However, misfolded proteins can overwhelm the proteostasis network. Protein misfolding is associated with numerous devastating neurodegenerative and cardiovascular disorders. Protein disaggregases can engage misfolded substrates and unfold them, promoting their return to proper fold and function. Disaggregases may serve as the final defense against collapse of the proteostasis network. Disaggregases play key roles in maintaining cellular health, and can even regulate beneficial amyloids in yeast. Amyloid is an exceptionally stable protein conformation that is implicated in numerous human diseases, and amyloid is considered to be otherwise intractable. I hypothesize that disaggregases play key roles in maintaining cellular health, but are vulnerable when the proteostasis network becomes overwhelmed in disease. Therefore, technologies that modulate disaggregase activity might be therapeutically useful. However, protein disaggregases are the least well characterized branch of the proteostasis network. I envision building a research program focused on developing a more comprehensive understanding of how disaggregases counter misfolding, both in health and under stress. I seek to elucidate how cells maintain proteostasis, how proteostasis fails, and how protein disaggregases might ultimately be applied to prevent or even reverse collapse of proteostasis. To further these goals, we will focus on three main themes over the next five years: (1) we will develop and apply new technologies to study and modulate the substrate-specificity of Hsp104, which regulates beneficial amyloid conformers in yeast. (2) We will characterize newly identified human amyloid disaggregases to better understand their normal roles in maintaining cellular health and how they might fail in disease. (3) We will apply new mass spectrometry techniques to study how protein-remodeling factors select specific substrates to target and how this varies under stress conditions. Our work will elucidate how disaggregases target specific substrates. Additionally, these finely-tuned disaggregases can be used as probes to test the hypothesis that misfolded species are toxic, and that restoration of proteins to their native folds and functions can reverse disease phenotypes. Ultimately we aim to apply the findings from these studies to develop new strategies to treat neurodegenerative disease. This is especially important because, despite intense efforts, there are no therapeutics available to treat protein-misfolding disorders.

抽象的： 蛋白质的正确折叠对于所有生命都至关重要，这使得异常的蛋白质折叠成为严重问题。这 蛋白质稳态（蛋白质稳态）网络是一个严格调控的系统，可确保蛋白质折叠 并根据需要进行降级。然而，错误折叠的蛋白质可能会破坏蛋白质稳态网络。蛋白质 错误折叠与许多毁灭性的神经退行性疾病和心血管疾病有关。蛋白质 解聚酶可以接合错误折叠的底物并将其展开，促进它们恢复到正确的折叠状态 功能。分解酶可能是防止蛋白质稳态网络崩溃的最后一道防线。 解聚酶在维持细胞健康方面发挥着关键作用，甚至可以调节有益的淀粉样蛋白 酵母。淀粉样蛋白是一种异常稳定的蛋白质构象，与许多人类疾病有关， 淀粉样蛋白被认为是难以治愈的。我假设分解在以下方面发挥着关键作用 维持细胞健康，但当蛋白质内稳态网络不堪重负时，就会变得脆弱 疾病。因此，调节解聚酶活性的技术可能在治疗上有用。然而， 蛋白质解聚酶是蛋白质稳态网络中特征最不明确的分支。我设想建立一个 研究计划的重点是更全面地了解分类如何对抗 错误折叠，无论是在健康还是在压力下。我试图阐明细胞如何维持蛋白质稳态，如何 蛋白质稳态失败，以及蛋白质解聚酶最终如何用于预防甚至逆转 蛋白质稳态崩溃。为了进一步实现这些目标，我们将在未来五年重点关注三个主题： (1)我们将开发和应用新技术来研究和调节Hsp104的底物特异性， 它调节酵母中有益的淀粉样蛋白构象异构体。 (2) 我们将表征新识别的人类 淀粉样蛋白解聚，以更好地了解它们在维持细胞健康中的正常作用以及它们如何 可能因疾病而失败。 (3)我们将应用新的质谱技术来研究蛋白质重构是如何进行的 因素选择特定的目标底物以及它在压力条件下如何变化。我们的工作将阐明 解聚集如何针对特定底物。此外，这些经过微调的解聚酶可用作 探针来检验错误折叠的物种是有毒的以及蛋白质恢复到其天然状态的假设 折叠和功能可以逆转疾病表型。最终我们的目标是应用这些研究的结果 开发治疗神经退行性疾病的新策略。这一点尤其重要，因为尽管 经过艰苦努力，目前还没有可用于治疗蛋白质错误折叠疾病的疗法。

项目成果

Probing the drivers of Staphylococcus aureus biofilm protein amyloidogenesis and disrupting biofilms with engineered protein disaggregases.

探究金黄色葡萄球菌生物膜蛋白淀粉样蛋白生成的驱动因素，并利用工程蛋白解聚酶破坏生物膜。

• 发表时间: 2023-08-31
• 影响因子: 6.4
• 作者: Howard, Matthew K;Miller, Karlie R;Sohn, Brian S;Ryan, Jeremy J;Xu, Andy;Jackrel, Meredith E
• 通讯作者: Jackrel, Meredith E

Drivers of Hsp104 potentiation revealed by scanning mutagenesis of the middle domain.

通过扫描中间结构域的诱变揭示了 Hsp104 增强的驱动因素。

• 发表时间: 2021
• 作者: Ryan, Jeremy J;Bao, Aaron;Bell, Braxton;Ling, Cendi;Jackrel, Meredith E
• 通讯作者: Jackrel, Meredith E

Molecular determinants and modifiers of Matrin-3 toxicity, condensate dynamics, and droplet morphology.

Matrin-3 毒性、冷凝物动力学和液滴形态的分子决定因素和调节剂。

• 发表时间: 2022-03-18
• 影响因子: 5.8
• 作者: Sprunger, Macy L;Lee, Ken;Sohn, Brian S;Jackrel, Meredith E
• 通讯作者: Jackrel, Meredith E

MANF stimulates autophagy and restores mitochondrial homeostasis to treat toxic proteinopathy.

MANF 刺激自噬并恢复线粒体稳态，以治疗毒性蛋白病。

• 发表时间: 2023-01-12
• 作者: Kim, Yeawon;Li, Chuang;Gu, Chenjian;Tycksen, Eric;Puri, Anuradhika;Pietka, Terri A;Sivapackiam, Jothilingam;Fang, Yili;Kidd, Kendrah;Park, Sun;Johnson, Bryce G;Kmoch, Stanislav;Duffield, Jeremy S;Bleyer, Anthony J;Jackrel, Meredith E;Ura
• 通讯作者: Ura

Therapeutic genetic variation revealed in diverse Hsp104 homologs.

多种 Hsp104 同源物中揭示了治疗性遗传变异。

• 发表时间: 2020-12-15
• 影响因子: 7.7
• 作者: March, Zachary M;Sweeney, Katelyn;Kim, Hanna;Yan, Xiaohui;Castellano, Laura M;Jackrel, Meredith E;Lin, JiaBei;Chuang, Edward;Gomes, Edward;Willicott, Corey W;Michalska, Karolina;Jedrzejczak, Robert P;Joachimiak, Andrzej;Caldwell, Kim A;Caldw
• 通讯作者: Caldw