CAM-KINASE II AT SYNAPTIC JUNCTIONS

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。我们正在使用我们的CaM-kinase II的3-D结构，该结构由单颗粒的3-D冷冻电镜重建产生，作为潜在的搜索模板，以识别突触连接处的CaM-kinase II分子（突触前和突触后元件）。NCMIR正在制作突触连接的高分辨率断层图，最初来自CA1锥体细胞上的突触，作为搜索的目标。在过去的一年里，已经制作了传统制备材料（例如，化学固定和嵌入环氧树脂）的CA1突触的单倾斜和双倾斜断层图，并评估了它们对该项目的适用性。下一步是使用高压冷冻和自由替代的材料。我们正在编写新的搜索算法来实现搜索功能的自动化。与此同时，我们正在评估用于初步可行性研究的直接可视化的能力。

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. We are using our 3-D structure of CaM-kinase II produced from 3-D cryo-EM reconstructions of single particles as a search template to potentially identify CaM-kinase II molecules at synaptic junctions (pre and post-synaptic elements). NCMIR is producing high resolution tomograms of synaptic junctions, initially from synapses on CA1 pyrnamidal cells, as the target for the search. During the past year, both single and double tilt tomograms of CA1 synapses from conventionally prepared material (e.g., chemically fixed and embedded in epoxy) have been produced and were evaluated for their suitability for this project. The next step is to use material that is high pressure frozen and free substituted. We are the process of writing new search algorithims to automate the search function. In the meanwhile, we are assessing the ability of direct visualization to be used for an initial feasibility studies.