REGULATION OF PHOSPHATIDYLINOSITOL METABOLISM

磷脂酰肌醇代谢对真核生物的生长至关重要 有机体。激动剂刺激的膜水解性 动物体内的多聚磷脂酰肌醇可刺激多种 细胞反应。一种酶，催化细胞内的第一步 合成的多聚磷脂酰肌醇是磷脂酰肌醇激酶。 这一应用将集中在磷脂酰肌醇的调节上 简单的真核生物酿酒酵母中的激酶。 磷脂酰肌醇激酶将被提纯到均一，并将被 在酶学、化学和物理方面具有特征 属性。纯化的酶将被重组成有明确定义的 研究单层磷脂微囊的活性调节作用。 将通过改变组成和环境来研究监管 小泡和可能的磷酸化和去磷酸化的 酵素。针对磷脂酰肌醇激酶的抗体将是 制备并用作探针以研究其调节作用 不同培养条件下细胞磷脂酰肌醇激酶的形成 条件。建议的研究应能让我们深入了解 多聚磷肌醇合成的调控。S。 酿酒酵母应该与高等真核生物相关。

Phosphatidylinositol metabolism is important to the growth of eukaryotic organisms. The agonist-stimulated hydrolysis of membrane polyphosphoinositides in animals result in the stimulation of various cellular responses. The enzyme that catalyzes the initial step in the synthesis of the polyphosphoinositides is phosphatidylinositol kinase. This application will focus on the regulation of phosphatidylinositol kinase in the simple eukaryote, Saccharomyces cerevisiae. Phosphatidylinositol kinase will be purified to homogeneity and will be characterized with respect to enzymological, chemical, and physical properties. The purified enzyme will be reconstituted into well defined unilamellar phospholipid vesicles to study the regulation of activity. Regulation will be studied by altering the composition and environment of the vesicles and by possible phosphorylation and dephosphorylation of the enzyme. Antibodies specific for phosphatidylinositol kinase will be prepared and used as a probe to study the regulation of phosphatidylinositol kinase formation in cells grown under various conditions. The proposed studies should provide insight into the regulation of polyphosphoinositide synthesis. The results found with S. cerevisiae should be relevant to higher eukaryotic organisms.