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

这一PI试图了解发育中的哺乳动物大脑的神经元是如何在大脑中适当定位的。在大脑发育期间，许多神经元从它们的出生地迁移，并受到环境中的线索的引导，建立起起作用的大脑的关键连接和结构。这项研究将使用生化、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但对其调控知之甚少的蛋白质。这项研究的一个主要焦点将是Ballif最近在这些小鼠大脑中的神经元定位蛋白上发现的新的酪氨酸磷酸化事件的特征。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在控制大脑发育的关键蛋白质中的作用。此外，这项研究将在发育过程中识别和量化数百种大脑蛋白质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可能需要数十年的工作。为了加速该领域其他发育神经生物学家的研究，所有已确定的磷酸化事件将提交到公共数据库，包括磷酸盐(http://www.phosphosite.org).在基因组革命之后，今天的本科生和研究生需要接受蛋白质组技术方面的培训。与佛蒙特州现有的基础设施和推广项目合作，Ballif博士将向佛蒙特州9所高等院校的150多名本科生教授蛋白质组学模块；将在研究项目中指导两名全日制研究生和10多名本科生暑期学生(包括少数族裔)；并将对蛋白质组学工作人员进行大规模定量蛋白质组学方面的培训，使其每年服务的许多研究人员都可以使用这项技术。

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.