THE EFFECT OF MOLECULAR TOXINS ON PROTEIN PHOSPHATASE 1 TARGETING

这个子项目是许多研究子项目中的一个 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。所列机构为 中心，不一定是研究者的机构。 分子毒素是一种天然产物，能有效抑制蛋白磷酸酶1（PP 1）的催化活性。这些天然抑制剂由许多生物体如海绵和蓝细菌产生，包括冈田酸、花萼菌素A、微囊藻毒素、节球藻素、互变霉素和fostriecin。我们希望全面了解这些分子毒素与PP 1紧密结合的结构和动力学机制。这将使我们能够表征由靶向和抑制蛋白与小分子毒素诱导的PP 1活性位点的细微分子差异。这些三维分子模型将使我们能够充分了解PP 1活性位点结构的细节。为了了解PP 1活性位点动力学及其在分子细节上靶向蛋白和抑制性配体（分子毒素）相互作用的微妙相互作用，我们将使用大量的生物化学和生物物理技术，特别是NMR光谱学和X射线晶体学，来阐明许多靶向蛋白和分子毒素本身的结构以及PP 1：靶向和PP 1：分子毒素复合物。PSP靶向蛋白，如神经元支架蛋白spinophilin，将PP 1靶向其作用点。这些相互作用的结构模型不存在，也不知道它们如何受到分子毒素相互作用的影响。因此，我们将使用多个PP 1：毒素和PP 1：靶向结构和动力学之间的差异和相似之处，以确定PP 1，一种普遍存在的丝氨酸/苏氨酸磷酸酶，是如何在细胞中有效和选择性地调节，使新的代理商，其目标是一个特定的群体的磷酸酶（即磷酸酶定位于多巴胺能神经元），可以开发。

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. Molecular toxins are natural products that potently inhibit the catalytic activity of protein phosphatase 1 (PP1). These natural inhibitors are created by numerous organisms such as marine sponges and cyanobacteria and include okadaic acid, calyculin A, microcystins, nodularins, tautomycin and fostriecin. We want to comprehensively understand the structural and dynamical mechanisms of the tight binding of these molecular toxins to PP1. This will enable us to characterize the subtle molecular differences of the PP1 active site induced by targeting and inhibitory proteins versus small molecule molecular toxins. These 3-dimensional molecular models will enable us to fully understand the details of the PP1 active site structure. To understand the PP1 active site dynamics, and its subtle interplay of targeting protein and inhibitory ligand (molecular toxin) interactions in molecular detail, we will use a large portfolio of biochemical and biophysical techniques, especially NMR spectroscopy and X-ray crystallography, to elucidate the structures of numerous targeting proteins and molecular toxins by themselves and as PP1:targeting and PP1:molecular toxin complexes. PSP targeting proteins, such as the neuronal scaffolding protein spinophilin, target PP1 to its point of action. No structural models for these interactions exist, nor is it known how they are affected by interactions with molecular toxins. Thus, we will use the differences and similarities between the multiple PP1:toxin and PP1:targeting structures and dynamics to determine how PP1, a ubiquitous ser/thr phosphatase, is potently and selectively regulated in the cell so that novel agents, which target a particular population of phosphatases (i.e. phosphatases localized in dopaminergic neurons), can be developed.