LINKING THE PROTEASOME ACTIVITY TO XPC BINDING PROTEIN HHR23A

将蛋白酶体活性与 XPC 结合蛋白 HHR23A 连接

• 批准号: 7721684
• 负责人: Kylie J. Walters
• 金额: $ 0.65万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721684
• 关键词: Binding; Binding Proteins; Bortezomib; Cells; Clinical; Complex; Computer Retrieval of Information on Scientific Projects Database; Event; Functional disorder; Funding; Goals; Grant; Immunocompetent; Institution; Lead; Life Cycle Stages; Link; Malignant Neoplasms; Methodology; Modeling; Monitor; Multiple Myeloma; NMR Spectroscopy; Neurodegenerative Disorders; Outcomes Research; Pathway interactions; Patients; Peptides; Polyubiquitin; Proteasome Inhibitor; Proteins; Research; Research Personnel; Resources; Source; Specificity; Staging; Structure; Ubiquitin; United States National Institutes of Health; Velcade; infancy; multicatalytic endopeptidase complex; protein degradation; receptor; receptor binding; success

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The ubiquitin proteasome pathway regulates an astounding array of cellular events and remains essential throughout the life cycle of a cell. It is therefore not surprising that its dysfunction is associated with ailments as threatening as cancer and neurodegenerative diseases. This pathway functions in two stages, substrate ubiquitylation, which culminates in the covalent attachment of polyubiquitin to a protein substrate, and proteasomal degradation, which results in the degradation of the protein substrate into immunocompetent peptides. The connection between these two events requires ubiquitin receptors. The goal of this project is to determine the mechanistic pathways connecting substrate ubiquitylation to proteasomal degradation. We use NMR spectroscopy to determine the structure of proteasomal and non-proteasomal ubiquitin receptors alone and complexed with polyubiquitin. These structures will provide fundamental information on how receptors recognize their ubiquitylated substrates. NMR is the ideal methodology for these studies, as we can readily monitor dynamic interactions between polyubiquitin and multiple binding partners. This attribute is a large asset, as ubiquitin receptors bind each other and such interactions likely provide an effective mechanism for shuttling ubiquitylated substrates to and within the proteasome. By using NMR, we will monitor polyubiquitin as it passed from one receptor to another, and thereby determine which interactions are mutually exclusive or prioritized. Such studies will lead to models of how ubiquitylated substrates are captured by their receptors and delivered to the proteasome. The outcome of this research could be used to reveal points of specificity in the ubiquitin-proteasome pathway. Notably, the clinical importance of this pathway has been demonstrated by the success of a specific proteasome inhibitor (bortezomib/Velcade) in treating patients with multiple myeloma. A mechanistic understanding of the ubiquitin proteasome pathway is in its infancy, as new ubiquitin receptors and proteasome components are still being revealed. We are part of a research team that has identified a new proteasomal ubiquitin receptor. As part of this proposal, we determine this protein¿s structure and define how it interacts with ubiquitin and other receptors.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以出现在其他 CRISP 条目中 列出的机构是。 对于中心来说，它不一定是研究者的机构。 泛素蛋白酶体途径调节一系列令人震惊的细胞事件，并且在细胞的整个生命周期中仍然至关重要，因此，其功能障碍与癌症和神经退行性疾病等威胁性疾病相关，这一点并不奇怪。泛素化，最终导致多聚泛素与蛋白质底物共价连接，以及蛋白酶体降解，导致蛋白质底物降解为免疫活性肽。这两个事件之间的联系需要泛素受体。该项目的目标是确定连接底物泛素化与蛋白酶体降解的机制途径。我们使用核磁共振波谱来确定单独的蛋白酶体和非蛋白酶体泛素受体以及与多聚泛素复合的结构。结构将提供有关受体如何识别其泛素化底物的基本信息，NMR 是这些研究的理想方法，因为我们可以轻松监测动态。多聚泛素和多个结合伙伴之间的相互作用是一个很大的优势，因为泛素受体相互结合，并且这种相互作用可能为泛素化底物穿梭于蛋白酶体和蛋白酶体内提供了有效的机制，我们将在多聚泛素通过时对其进行监测。从一种受体到另一种受体，并确定哪些相互作用是相互排斥或优先的，从而此类研究将产生泛素化底物如何被其受体捕获并传递至受体的模型。这项研究的结果可用于揭示泛素-蛋白酶体途径的特异性点。值得注意的是，特定蛋白酶体抑制剂（硼替佐米/万卡德）在治疗患有乳腺癌的患者中的成功证明了该途径的临床重要性。对泛素蛋白酶体途径的机制了解还处于起步阶段，因为新的泛素受体和蛋白酶体成分仍在被发现。是一个研究小组的成员，该研究小组已经确定了一种新的蛋白酶体泛素受体作为该提案的一部分，我们确定了这种蛋白质。的结构并定义它如何与泛素和其他受体相互作用。