Phosphotyrosine-dependent regulatory mechanisms of mammalian brain development: A large-scale phosphoproteomic and biochemical study.

本PI旨在了解发育中的哺乳动物大脑的神经元如何在大脑中适当定位。在大脑发育过程中，许多神经元从它们出生的地方迁移，并在环境中的线索的引导下形成功能性大脑的关键连接和结构。这项研究将使用生物化学，细胞生物学和新开发的蛋白质组学方法来研究已知对神经元定位至关重要的蛋白质，但其调控机制知之甚少。这项研究的一个主要重点将是新的酪氨酸磷酸化事件的特征，最近确定的Ballif对这些神经元定位蛋白从小鼠大脑。酪氨酸磷酸化是可逆调节蛋白质和改变细胞命运的最动态机制之一。预计这项研究将明确靶向酪氨酸磷酸化事件对控制大脑发育的关键蛋白质的作用。此外，这项研究将在整个发育过程中识别和量化数百种脑蛋白上的数千种酪氨酸磷酸化事件，在几年内完成使用传统方法需要几十年的工作。为了加速该领域其他发育神经生物学家的研究，所有鉴定的磷酸化事件将提交给包括Phosphosite（http：//www.example.com）在内的公共数据库。www.phosphosite.org在基因组学革命之后，今天的本科生和研究生需要获得蛋白质组学技术的培训。Ballif博士将与佛蒙特州的基础设施和外展项目合作，向佛蒙特州9所高等院校的150多名本科生教授蛋白质组学模块;将指导两名全日制研究生和10多名本科生的研究项目（包括少数民族）;并将对蛋白质组学的工作人员进行大规模的培训，大规模定量蛋白质组学使这项技术可用于它每年服务的许多研究人员。

This PI seeks to understand how neurons of the developing mammalian brain are appropriately positioned in the brain. During brain development many neurons migrate from their places of birth and are guided by cues in their environment to make the critical connections and structures of a functioning brain. This study will use biochemical, cell biological and newly developed proteomic approaches to study proteins that are known to be essential for neuronal positioning, but whose regulation is poorly understood. A major focus of the study will be the characterization of novel tyrosine phosphorylation events recently identified by Ballif on these neuronal-positioning proteins from mouse brain. Tyrosine phosphorylation is one of the most dynamic mechanisms reversibly regulating proteins and altering cell fate. It is anticipated that this study will make clear the role of targeted tyrosine phosphorylation events on critical proteins governing brain development. Additionally, this study will identify and quantify across development many thousands of tyrosine phosphorylation events on hundreds of brain proteins, accomplishing in a few years what would have required decades using traditional methods. To accelerate the research of other developmental neurobiolists in the field, all identified phosphorylation events will be submitted to public databases including Phosphosite (http://www.phosphosite.org). On the heels of the genomic revolution today's undergraduate and graduate students need access to training in proteomic technology. In collaboration with established infrastructure and outreach programs in Vermont, Dr. Ballif will teach a proteomic module to over 150 undergraduates at nine institutions of higher learning in Vermont; will mentor in research projects two full time graduate students and more than ten undergraduate summer students (including minorities); and will train the proteomics staff in large-scale quantitative proteomics making this technology available to the many investigators it serves each year.