STRUCTURE DETERMINATION OF THE FERM DOMAIN OF PYK2 IN COMPLEX WITH THE

PYK2 复合物的 FERM 结构域的结构测定

• 批准号: 8363541
• 负责人: JOSEPH SCHLESSINGER
• 金额: $ 0.69万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363541
• 关键词: Binding; Blood capillaries; C-terminal; Collaborations; Complex; Data Collection; Data Set; Funding; Grant; National Center for Research Resources; Nature; PTK2B gene; Principal Investigator; Property; Research; Research Infrastructure; Resolution; Resources; Signal Transduction; Source; Structure; Techniques; Temperature; Testing; Time; United States National Institutes of Health; capillary; cost; improved; pressure; Binding; Blood capillaries; C-terminal; Collaborations; Complex; Data Collection; Data Set; Funding; Grant; National Center for Research Resources; Nature; PTK2B gene; Principal Investigator; Property; Research; Research Infrastructure; Resolution; Resources; Signal Transduction; Source; Structure; Techniques; Temperature; Testing; Time; United States National Institutes of Health; capillary; cost; improved; pressure

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. Structure determination of the Ferm domain of Pyk2 in complex with the C-terminal domain of Nir2 We have obtained crystals of Ferm domain of Pyk2 in complex with the C-terminal domain of Nir2 that can elucidate how the C-terminal domain of Nir2 binds to the Ferm domain of Pyk2 and regulates Pyk2 activation and signaling. Intense crystal optimization of the complex as well as cryoprotectant search has been pursued yielding the datasets of resolution limit of 3.5 ¿ at X29 at Brookhaven. The limit of diffraction ranges of the crystals turned out to be caused by the cryoprotectants since the room temperature diffraction in the capillary gave higher resolution. However, the exposure of crystals to the beam at room temperature rapidly damaged crystals resulting that the data collection at room temperature is unlikely. Due to the nature of the complex crystals, we need newly developed technique, the high-pressure cooling. In collaboration with Dr. Sol M. Gruner and Dr. Chae Un Kim at Cornell, we are attempting to improve the resolution of our crystals using their newly developed high-pressure cryocooling approach. We are therefore requesting one (or two) day beam time at CHESS to test the effects of high-pressure cryocooling on the diffraction properties of these crystals, and if successful, collecting full native datasets.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 与NIR2的C末端结构域中Pyk2的FERM结构域的结构测定 我们已经在与NIR2的C末端结构域中获得了PYK2的FERM域的晶体，这些结构域可以阐明NIR2的C末端结构域如何与PYK2的FERM结合并调节PYK2激活和信号传导。在Brookhaven的X29上，已追求了复合物和冷冻保护剂搜索的强烈晶体优化，并产生了3.5€的分辨率限制的数据集。晶体的衍射范围的极限原来是由冷冻保护剂引起的，因为毛细管中的室温衍射给出了更高的分辨率。但是，在室温下，晶体暴露于横梁迅速损坏，导致不太可能在室温下收集数据。由于复杂晶体的性质，我们需要新开发的技术，即高压冷却。与Sol M. Gruner博士和Cornell的Chae Un Kim博士合作，我们试图使用其新开发的高压冷冻方法来改善晶体的分辨率。因此，我们要求在国际象棋时在国际象棋上进行一（或两次）的光束时间，以测试高压冷冻冷却对这些晶体衍射特性的影响，如果成功的话，收集了完整的本机数据集。