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

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH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 中心，不一定是研究者的机构。 Pyk 2的Ferm结构域与Nir 2的C末端结构域复合物的结构确定 我们已经获得了Pyk 2的Ferm结构域与Nir 2的C-末端结构域复合的晶体，这可以阐明Nir 2的C-末端结构域如何结合Pyk 2的Ferm结构域并调节Pyk 2活化和信号传导。对复合物进行了密集的晶体优化，并进行了冷冻保护剂搜索，在Brookhaven的X29处获得了分辨率极限为3.5“的数据集。晶体衍射范围的限制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。然而，晶体在室温下暴露于光束会迅速损坏晶体，导致在室温下不可能收集数据。由于复杂晶体的性质，我们需要新开发的技术，高压冷却。与Sol M博士合作。Gruner和康奈尔大学的Chae Un Kim博士，我们正试图使用他们新开发的高压低温冷却方法来提高晶体的分辨率。因此，我们要求在CHESS进行一天（或两天）的光束时间，以测试高压低温冷却对这些晶体衍射特性的影响，如果成功，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.