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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 PYK2的FERM结构域与Nir2的C-末端结构域的络合物的结构测定 我们已经获得了PYK2的FERM结构域与Nir2的C-末端结构域的络合物的晶体，可以阐明Nir2的C-末端结构域是如何与PYK2的FERM结构域结合并调节PYK2的激活和信号转导的。对该络合物进行了密集的晶体优化以及低温保护剂的搜索，在布鲁克海文获得了分辨率极限为3.5°的数据集。由于毛细管室温衍射具有较高的分辨率，晶体的极限衍射范围是由低温保护剂引起的。然而，晶体在室温下暴露在光束下会迅速损坏晶体，因此在室温下收集数据是不可能的。由于复杂晶体的性质，我们需要新开发的技术--高压冷却。在康奈尔大学的Sol M.Gruner博士和Chae Un Kim博士的合作下，我们正在尝试使用他们新开发的高压冷冻方法来提高我们晶体的分辨率。因此，我们在CHESS请求一天(或两天)的束流时间，以测试高压冷冻对这些晶体的衍射特性的影响，如果成功，将收集完整的本地数据集。

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. 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.