Hybrid Phospholipid-Phosphoinositides

描述（由申请人提供）：结合蛋白和脂质代谢酶对磷酸肌肽的识别涉及与头基团和二酰基甘油主链的特定相互作用，这种相互作用因蛋白质而异。当将荧光探针引入肌醇头组时，结合和代谢被严重减弱或完全取消。当将荧光探针引入酰基时，许多衍生物不能显示出足够的Km和Vmax值，作为脂质激酶和磷酸酶的底物，也不能与同源的pip结合蛋白结合。最近，埃施朗生物科学公司开发了一种基于四碳苏糖醇骨架的磷脂-磷酸苷杂合体的新方法。这些新材料，被称为Pea-PIPs TM，具有磷脂酰乙醇胺头基团，允许生化或化学探针附着在与磷酸肌苷头基团和两个酰基链正交的空间位置。在i期授权中，我们将首先合成所有8个Pea-PIPns TM的完整曲目。其次，我们将制备两个具有四种不同亲水性连接臂的Pea-PIPs TM，以优化探针头基团距离。第三，我们将确定使用Pea- PIPs TM作为脂质激酶、脂质磷酸酶和脂质结合蛋白的体外底物的可行性。

DESCRIPTION (provided by applicant): Phosphoinositide recognition by binding proteins and lipid-metabolizing enzymes involves specific interactions with the head group and diacylglycerol backbone, which vary significantly from protein to protein. When a fluorescent probe is introduced into the inositol head group, binding and metabolism are severely attenuated or abrogated entirely. When a fluorescent probe is introduced into the acyl groups, many of the resulting derivatives fail to show adequate Km and Vmax values as substrates for lipid kinases and phosphatases as well as binding to cognate PIP-binding proteins. Recently, Echelon Biosciences has developed a new approach to make phospholipid-phophoinositide hybrids based on a four-carbon threitol backbone. These new materials, known as Pea-PIPs TM, have a phosphatidylethanolamine head group that permits biochemical or chemical probes to be attached in a position spatially orthogonal to the phosphoinositide head group and the two acyl chains. In this Phase i grant, we will first synthesize a full repertoire of all eight Pea-PIPns TM. Second, we will prepare two Pea-PIPs TM with four different hydrophilic linker arms to optimize probe-head group distances. Third, we will establish the feasibility of using Pea- PIPs TM as in vitro substrates for lipid kinases, lipid phosphatases, and lipid binding proteins.