HSP90 CONFORMATIONAL REQUIREMENTS FOR KINASE MATURATION

HSP90 激酶成熟的构象要求

• 批准号: 8168638
• 负责人: DANIEL N BOLON
• 金额: $ 0.54万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168638
• 关键词: Binding; Biology; C-terminal; Computer Retrieval of Information on Scientific Projects Database; Data; Engineering; Eukaryota; Funding; Grant; Institution; Medicine; Modeling; Molecular; Molecular Chaperones; Molecular Conformation; N Domain; N-terminal; Oncogenic; Phosphotransferases; Process; Research; Research Personnel; Resources; Roentgen Rays; Source; Structure; United States National Institutes of Health; anti-cancer therapeutic; design; dimer; in vivo; inhibitor/antagonist; insight; research study

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. Hsp90 is an essential chaperone in eukaryotes that is involved in the maturation of numerous kinases. Because numerous oncogenic kinases rely on Hsp90 for their function, Hsp90 has emerged as a promising target for anti-cancer therapeutics. The design of effective Hsp90 inhibitors would benefit from a molecular understanding of Hsp90 mechanism. Recently, two structures of Hsp90 have been determined both with the C-terminal domain forming a stable dimer. In one structure the N-terminal domain is also dimerized forming a closed state, and in the other structure the N-terminal domain is separated by a large distance (>60 Angstroms). Small-angle X-ray scattering (SAXS) experiments are consistent with a model where ATP-binding shifts Hsp90 from the open to the closed conformation. To determine if the open conformation is involved in kinase maturation, we have engineered Hsp90 with a coiled-coil to hold the N-domain in the closed conformation in the absence of ATP binding. SAXS experiments at APS would provide critical data to determine the conformational states of our engineered Hsp90 molecules both in the presence and absence of ATP. Combined with in vivo studies currently underway to analyze kinase maturation, the SAXS data will provide important insights into a fundamental process in biology and medicine.

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。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 Hsp90 是真核生物中重要的伴侣，参与多种激酶的成熟。由于许多致癌激酶的功能依赖于 Hsp90，因此 Hsp90 已成为抗癌治疗的一个有前景的靶标。有效 Hsp90 抑制剂的设计将受益于对 Hsp90 机制的分子理解。最近，Hsp90 的两种结构已被确定，两者的 C 端结构域均形成稳定的二聚体。在一种结构中，N 端结构域也二聚化，形成闭合状态，而在另一种结构中，N 端结构域相隔较远的距离（> 60 埃）。小角 X 射线散射 (SAXS) 实验与 ATP 结合将 Hsp90 从开放构象转变为闭合构象的模型一致。为了确定开放构象是否参与激酶成熟，我们设计了具有卷曲螺旋的 Hsp90，以在没有 ATP 结合的情况下将 N 结构域保持在闭合构象。 APS 的 SAXS 实验将提供关键数据，以确定我们工程化的 Hsp90 分子在存在和不存在 ATP 的情况下的构象状态。结合目前正在进行的分析激酶成熟的体内研究，SAXS 数据将为生物学和医学的基本过程提供重要的见解。