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

调节动物极性和纺锤体方向的分子机制 - 购买质谱仪的行政补充

• 批准号: 9893401
• 负责人: Kenneth E Prehoda
• 金额: $ 7.03万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9893401
• 关键词: Administrative Supplement; Animals; Biological Models; Cell Polarity; Complement; Complex; Funding; Laboratories; Mass Spectrum Analysis; Methods; Molecular; Post-Translational Protein Processing; Reagent; Reproducibility; Research; Resolution; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; System; Techniques; Work; atypical protein kinase C; gel electrophoresis; improved; instrument; mass spectrometer; neuroblast; parent grant; polarized cell; programs; protein complex; reconstitution

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.

项目摘要!