PROTEINS IMPORTANT FOR PROGRAMMED CELL DEATH AND FOR GENE-LINKED BLINDNESS

对于程序性细胞死亡和基因相关失明很重要的蛋白质

• 批准号: 8362355
• 负责人: CHARLES EIGENBROT
• 金额: $ 0.14万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362355
• 关键词: Age related macular degeneration; Amino Acids; Apoptosis; Apoptosis Inhibitor; Apoptotic; Binding; Blindness; C-terminal; Cell Death; DNA Sequence Rearrangement; Data; Distant; Event; Funding; Genes; Grant; Homologous Gene; Human; Immunoglobulin Fragments; Inhibition of Apoptosis; Insulin-Like Growth-Factor-Binding Proteins; Learning; Link; Location; N-terminal; National Center for Research Resources; Peptide Hydrolases; Population; Predisposition; Principal Investigator; Protease Domain; Protein Binding Domain; Proteins; Radiation; Research; Research Infrastructure; Resources; Role; Source; Ubiquitination; United States National Institutes of Health; Variant; blocking factor; cancer therapy; cost; dimer; molecular shape; monomer; overexpression; small molecule; structural biology; ubiquitin-protein ligase

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Inhibitors of apoptosis (IAPs) are anti-apoptotic factors that block cell death. IAPs contain protein-protein interaction domains and a C-terminal RING domain that confers E3 ubiquitin ligase activity. Binding of small molecule antagonists to cellular IAPs promotes cIAP auto-ubiquitination and subsequent proteasomal degradation, thereby releasing cIAP inhibition of apoptosis?a situation that may render some cancer therapies more effective. How the distant antagonist binding event influences the E3 ligase activity of the physically distinct RING domain is unclear. Our preliminary data support a hypothesis where antagonist binding leads to a structural rearrangement that permits formation of active RING-RING dimers. Moreover, antagonists can also promote a cIAP1 population of intermediate size; larger than monomer, yet smaller than a dimer. This form of cIAP1 is the dominant species in the presence of a particular monovalent antagonist and may represent the ?open?, but un-dimerized, form of the protein. We seek SAXS data from cIAP1 both in the absence and presence of small molecule antagonists. Htra1 is a multi-domain protease. A variant human Htra1 gene promotes overexpression and increases susceptibility to age-related macular degeneration. Htra1 is composed of four domains for which molecular shapes are known by homology and its protease activity requires formation of a non-covalent homotrimer of 1440 amino acids. The functional roles of the non-protease domains are not clear. The N-terminal domain composed of the IGFBP-rP and Kazal domains is absent in baterial homologues, suggesting a unique regulatory function. Our objective is to learn the domain locations in the trimer, potential domain interactions and domain oligomerization status. In addition, we have anti-Htra1 antibody fragments which may aid in SAXS analysis.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 细胞凋亡抑制剂 (IAP) 是阻止细胞死亡的抗细胞凋亡因子。 IAP 包含蛋白质-蛋白质相互作用结构域和赋予 E3 泛素连接酶活性的 C 端 RING 结构域。 小分子拮抗剂与细胞 IAP 的结合促进 cIAP 自动泛素化和随后的蛋白酶体降解，从而释放 cIAP 对细胞凋亡的抑制——这种情况可能会使一些癌症治疗更加有效。 远距离拮抗剂结合事件如何影响物理上不同的 RING 结构域的 E3 连接酶活性尚不清楚。 我们的初步数据支持这样的假设：拮抗剂结合导致结构重排，从而允许形成活性环-环二聚体。 此外，拮抗剂还可以促进中等规模的 cIAP1 群体；比单体大，但比二聚体小。 这种形式的 cIAP1 在特定单价拮抗剂存在下是主要种类，并且可能代表蛋白质的“开放”但非二聚化形式。 我们在不存在和存在小分子拮抗剂的情况下寻求 cIAP1 的 SAXS 数据。 Htra1 是一种多结构域蛋白酶。人类 Htra1 基因变异会促进过度表达并增加对年龄相关性黄斑变性的易感性。 Htra1 由四个结构域组成，其分子形状通过同源性已知，其蛋白酶活性需要形成 1440 个氨基酸的非共价同源三聚体。非蛋白酶结构域的功能作用尚不清楚。 由 IGFBP-rP 和 Kazal 结构域组成的 N 端结构域在细菌同源物中不存在，表明其具有独特的调节功能。我们的目标是了解三聚体中的结构域位置、潜在的结构域相互作用和结构域寡聚状态。此外，我们还有抗 Htra1 抗体片段，可能有助于 SAXS 分析。