ER and Post-ER Quality Control of Integral Membrane Proteins

完整膜蛋白的 ER 和 ER 后质量控制

• 批准号: 10798491
• 负责人: JEFFREY L. BRODSKY
• 金额: $ 1.76万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10798491
• 关键词: 26S proteasome; Biochemical; Cell secretion; Cells; Cytoplasm; Cytosol; Degradation Pathway; Development; Disease; Endoplasmic Reticulum; Eukaryota; Event; Foundations; Genetic Screening; Goals; Health; Integral Membrane Protein; Link; Membrane; Membrane Proteins; Modeling; Molecular; Molecular Chaperones; Names; Organelles; Paper; Pathway interactions; Peptide Hydrolases; Play; Process; Protein Biosynthesis; Publications; Publishing; Quality Control; Research; Resistance; Role; Sorting; Stress; Technology; Translation Process; Ubiquitin; Ubiquitination; disease-causing mutation; experimental study; extracellular; human disease; in vitro Assay; member; method development; misfolded protein; polypeptide; posters; programs; protein aggregation; protein folding; proteotoxicity; tool

Approximately one-third of all newly synthesized proteins in eukaryotes enter the endoplasmic reticulum (ER). Once associated with this compartment, these nascent polypeptides are post- translationally processed, acquire their native confirmations, oligomerize, and are sorted for extracellular secretion or delivery to other organelles. However, many disease-causing mutations compromise protein folding and maturation, which in turn can generate aggregation- prone species. To off-set the catastrophic effects that accompany the accumulation of protein aggregates, misfolded protein substrates are: (i) selected by molecular chaperones associated with the ER, (ii) modified with ubiquitin, (iii) delivered to the cytoplasm via a process known as retrotranslocation, and (iv) degraded by the 26S proteasome. Brodsky and colleagues named this pathway ER associated degradation (ERAD), and over the past 21 years many of the molecular mechanisms underlying this sequence of events were defined in the Brodsky lab. To date, ~80 human diseases are linked to the ERAD pathway and >1,200 publications have been authored on various aspects of this pathway. Ongoing efforts are defining the pathophysiological foundation of several ERAD-related disorders. In parallel, members of the Brodsky lab have revealed how key components orchestrate each step during ERAD. In the past 5 years, the lab has published 64 papers, and tools and technologies were developed that provide an unprecedented view of the mechanisms that lead to the selection, ubiquitination, retrotranslocation, and degradation of diverse substrates. Nevertheless, recent discoveries dictate that more challenging research directions are pursued: By necessity, these next efforts will require additional method development and a pursuit of longer-term goals. Specific questions that the research program will address include: What biochemical features define an ERAD substrate? Which factors are sufficient to drive the retrotranslocation of ERAD substrates, some of which are aggregation-prone? Do ER-associated proteases function in tandem with the 26S proteasome to destroy substrates that are stably integrated into the ER membrane, and thus might be retrotranslocation resistant? And, how are retrotranslocated membrane proteins— which can reside in the cytosol after being liberated from the ER—retained in a soluble state? Answers to these questions, which lie at the core of research in the field, will significantly advance an understanding of how cellular health is maintained in the face of proteotoxic stress as well as how ERAD-associated diseases arise and might be rectified.

真核生物中大约三分之一新合成的蛋白质进入内质 一旦与该区室结合，这些新生的多肽就被后置。 翻译处理，获取其本机确认，寡聚，并分类 然而，细胞外分泌或输送到其他细胞器却引起许多疾病。 突变会损害蛋白质折叠和成熟，进而产生聚集 抵消蛋白质积累带来的灾难性影响。 聚集体，错误折叠的蛋白质底物是：（i）由相关的分子伴侣选择 与 ER 一起，(ii) 用泛素修饰，(iii) 通过称为 逆转录转位，(iv) 被 26S 蛋白酶体降解，并命名为同事。 该途径与 ER 相关降解 (ERAD) 相关，在过去 21 年中，许多 布罗德斯基实验室定义了这一系列事件背后的分子机制。 迄今为止，约有 80 种人类疾病与 ERAD 通路相关，并且已发表超过 1,200 篇出版物 关于该途径各个方面的持续努力正在定义。 同时，几种 ERAD 相关疾病的病理生理学基础。 Brodsky 实验室揭示了 ERAD 期间关键组件如何协调每个步骤。 过去5年，该实验室发表了64篇论文，并开发了工具和技术 对导致选择、泛素化、 然而，最近的发现。 决定了要追求更具挑战性的研究方向：必然地，这些下一步的努力 将需要额外的方法开发和对长期目标的追求。 该研究计划将解决的问题包括： 哪些生化特征定义了 ERAD 底物哪些因素足以驱动 ERAD 底物的逆转位， 其中一些易于聚集？ ER 相关蛋白酶是否与 26S 蛋白酶体破坏稳定整合到 ER 膜中的底物，以及 那么逆转位膜蛋白是如何抵抗逆转位的呢？ 从内质网中释放出来后，它可以驻留在细胞质中——保留在可溶状态吗？ 这些问题的答案是该领域研究的核心，将显着 了解面对蛋白毒性应激如何维持细胞健康进步 以及 ERAD 相关疾病如何产生以及如何纠正。

项目成果

The expression system influences stability, maturation efficiency, and oligomeric properties of the potassium-chloride co-transporter KCC2.

表达系统影响氯化钾协同转运蛋白 KCC2 的稳定性、成熟效率和寡聚特性。

• 发表时间: 2024-03
• 影响因子: 4.2
• 作者: Kok, Morgan;Hartnett;Happe, Cassandra L;MacDonald, Matthew L;Aizenman, Elias;Brodsky, Jeffrey L
• 通讯作者: Brodsky, Jeffrey L

Ubiquitination of disease-causing CFTR variants in a microsome-based assay.

基于微粒体的检测中致病 CFTR 变异的泛素化。

• 发表时间: 2020
• 影响因子: 2.9
• 作者: Estabrooks, Samuel K;Brodsky, Jeffrey L
• 通讯作者: Brodsky, Jeffrey L

Hsp40s play distinct roles during the initial stages of apolipoprotein B biogenesis.

Hsp40 在载脂蛋白 B 生物发生的初始阶段发挥着独特的作用。

• 发表时间: 2022-02-01
• 影响因子: 3.3
• 作者: Kumari, Deepa;Fisher, Edward A;Brodsky, Jeffrey L
• 通讯作者: Brodsky, Jeffrey L

Fundamental and translational research in Cystic Fibrosis - why we still need it.

囊性纤维化的基础和转化研究 - 为什么我们仍然需要它。

• 发表时间: 2023-03
• 作者: Farinha, Carlos M;Brodsky, Jeffrey L;Pedemonte, Nicoletta
• 通讯作者: Pedemonte, Nicoletta

Excess dietary sodium partially restores salt and water homeostasis caused by loss of the endoplasmic reticulum molecular chaperone, GRP170, in the mouse nephron.

膳食中过量的钠可以部分恢复小鼠肾单位中因内质网分子伴侣 GRP170 丢失而引起的盐和水稳态。

• 发表时间: 2024-01-14
• 作者: Porter, Aidan;Vorndran, Hannah E;Marciszyn, Allison;Mutchler, Stephanie M;Subramanya, Arohan R;Kleyman, Thomas R;Hendershot, Linda M;Brodsky, Jeffrey L;Buck, Teresa M
• 通讯作者: Buck, Teresa M