Molecular mechanisms that regulate polarity and spindle orientation in animals - admin supplement to purchase mass spectrometer

项目摘要！ 申请资金用于购买Bruker Micro MicroTM SMART LS MALDI-TOF质谱仪 用于验证我们生产的试剂的分光计。该仪器将由我的 研究小组和我的同事布拉德诺伦谁提交了一个交叉引用 应用程序.在R35 GM 127092中，本补充申请的母基金，我的实验室 提出以神经母细胞为模型，揭示动物细胞极化的机制 系统一个关键的目标是使用重组纯化的艾德Par复合蛋白来了解 调节极性的机制。虽然我们通常使用凝胶电泳作为 用于确定我们试剂的同一性和纯度的方法，我们系统的几个特点 使质谱分析法成为该标准技术的重要补充。一是 Par复合物的一种成分，非典型蛋白激酶C，通过凝胶异常迁移 由于必需的翻译后修饰，电泳。第二，我们的工作 需要制备各种修饰的艾德形式的Par复合物蛋白（例如截短物）， 仪器将使我们能够用高分辨率的质量信息来验证它们。我们预计 该工具提供的额外信息将具有重要和持续的意义， 通过提高关键数据的可靠性和可重复性， 试剂

Project abstract! Funding is requested for the purchase of a Bruker Microflex SMART LS MALDI-TOF mass spectrometer for verifying the reagents we produce. The instrument will be used by my research group and by my colleague Brad Nolen who has submitted a cross-referenced application. In R35GM127092, the parent grant for this supplement application, my laboratory proposes to uncover the mechanism of animal cell polarization using the neuroblast as a model system. A key aim is to use reconstituted purified Par complex proteins to understand the mechanism by which polarity is regulated. While we routinely use gel electrophoresis as a method for determining the identity and purity of our reagents, several features of our system make mass spectrometry an important complement to this standard technique. First, a component of the Par complex, atypical Protein Kinase C, migrates abnormally by gel electrophoresis because of essential post translational modifications. Second, our work requires making various modified forms of Par complex proteins (e.g. truncations) and this instrument will allow us to verify them with high resolution mass information. We expect that the additional information provided by this instrument will have a significant and sustained positive impact on our research program by increasing the reliability and reproducibility of key reagents.