Functional Anatomy of Mammalian Phosphatidylinositol Transfer Proteins

 描述（由申请人提供）：本研究的目的是对研究不足的一类蛋白质进行详细分析：哺乳动物磷脂酰肌醇/磷脂酰胆碱转移蛋白（PITP）。哺乳动物PITP的功能和机制仍有待阐明。该研究计划旨在确定特定哺乳动物PITP亚型-1类PITP的功能机制。该建议是建立在我们的创建和表征的PITP基因敲除小鼠和一个等位基因系列的小鼠与分级缺陷的功能，这种蛋白质。pitp 0/0 小鼠及其衍生物是有吸引力的疾病模型，因为它们出生时是活的，但出生后表现出强大的表型。这些表型允许收集确定的基于细胞的试剂用于分析。使用这些独特的动物和细胞模型作为主要的分析对象，我们将进行三条线的调查。 首先，我们将使用生物化学和生物物理学的方法来破译1类PITP如何在单分子水平上发挥作用。 其次，我们将使用复杂的定量成像和分析测定来确定1类PITP如何在功能上参与信号传导受体酪氨酸激酶，如表皮生长因子受体和血小板衍生生长因子受体。这些受体的适当活性是适当的细胞生长控制和形态发生反应所必需的。神经错乱会导致癌症和神经缺陷。 第三，我们将利用我们组装的强大筛选平台，以鉴定和验证对1类PITP具有高特异性的小分子抑制剂。这样的试剂将作为用于细胞中磷酸肌醇信号传导的手术操作的工具化合物是有价值的，并且具有鉴定用于针对癌症或神经缺陷的治疗的新药的先导化合物的潜力。 PITP在调节与不同细胞过程相互作用的信号转导途径中发挥核心作用。然而，根本的机制是不理解的，因为这些还没有被调查。Bankaitis实验室独特地准备解决1类PITP功能的机制问题，因为它已经开发了独特的实验系统进行分析。

 DESCRIPTION (provided by applicant): The objective of this research is to undertake a detailed analysis of an under-investigated class of proteins: the mammalian phosphatidylinositol/phosphatidylcholine transfer proteins (PITPs). The functions and mechanisms of mammalian PITP function remains to be elucidated. The research plan is designed to identify mechanisms of function of specific mammalian PITP isoforms - the Class 1 PITPs. This proposal is founded on our creation and characterization of a PITP knockout mouse and an allelic series of mice with graded defects in function of this protein. The pitp0/0 mouse and its derivatives are attractive disease models in that these are born alive, but manifest powerful phenotypes after birth. These phenotypes permit collection of defined cell-based reagents for analysis. Using these unique animal and cellular models as primary analytical subjects, we will undertake three lines of investigation. First, we will use sophisticaed biochemical and biophysical approaches to decipher how Class 1 PITPs function at the level of single molecules. Second, we will use sophisticated and quantitative imaging and analytical assays to determine how Class 1 PITPs functionally engages signaling receptor tyrosine kinases such as epidermal growth factor receptor and platelet-derived growth factor receptor. The appropriate activity of these receptors is required for proper cell growth control and morphogen responses. Derangements lead to cancer and neurological deficits. Third, we will exploit a powerful screening platform we assembled for the purpose of identifying and validating small molecule inhibitors with high specificity for Class 1 PITPs. Such reagents would be of value as tool compounds for surgical manipulation of phosphoinositide signaling in cells, and hold the potential of identifying lead compounds for new pharmaceuticals directed at treatment of cancer or neurological deficits. PITPs play central roles in regulating signal transduction pathways that interface with diverse cellular processes. Yet, the underlying mechanisms are not understood because these have not been investigated. The Bankaitis laboratory is uniquely poised to address questions of mechanism of Class 1 PITP function as it has developed unique experimental systems for analysis.