CAM-KINASE II AT SYNAPTIC JUNCTIONS

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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.