GENETICS OF FLAVIN DEHYDROGENASE-MEMBRANE INTERACTIONS

这项研究的目的是确定黄化如何 沙门氏菌中脱氢酶与膜的相互作用 鼠伤寒杆菌。这项工作将主要围绕两个主题展开： 1.对看跌期权操纵者的监管。PUT操作员有一个独特的 似乎是由puta基因介导的控制机制 产品，脯氨酸氧化酶。当外源脯氨酸可用时， 脯氨酸氧化酶结合到膜上，在那里它发挥着 黄素脱氢酶；但在没有脯氨酸的情况下，它 积聚在细胞质中，在那里它充当自体 抑制者。确定Pro氧化酶是如何控制自身的 表达可能有助于我们理解其他膜结合是如何 调节蛋白起作用。此外，的监管属性 PUTA突变为细胞膜提供了基因选择 相互作用突变体。 2.黄素脱氢酶-膜相互作用。剥离 脱氢酶必须与膜结合才能 与电子传输链正确地相互作用。然而，它 不知道它们是如何与膜相互作用的 它们相互作用的电子传输链的组成部分 直接去吧。脯氨酸氧化酶相互作用的遗传学研究 为理解膜提供了一个模型系统 蛋白质在体内如何与细胞膜相互作用。蛋白质- 膜所需的蛋白质和蛋白质-脂相互作用 关联性可以通过分离符合以下条件的突变来确定 防止脯氨酸氧化酶与细胞膜结合。遗传 对这些突变体的生化分析将表明 膜组件(S)与黄素直接相互作用 脱氢酶以及需要什么样的相互作用 黄素脱氢酶与膜电子之间的关系 运输链。这样的突变体应该能让我们深入了解 外周膜蛋白如何存在的一般生物学问题 与膜相互作用。

The objective of this study is to determine how flavin dehydrogenases interact with the membrane in Salmonella typhimurium. This work will focus on two main topics: 1. Regulation of the put operon. The put operon has a unique control mechanism that seems to be mediated by the putA gene product, proline oxidase. When exogenous proline is available, proline oxidase binds to the membrane where it functions as a flavin dehydrogenase; but in the absence of proline, it accumulates in the cytoplasm where it acts as an autogenous repressor. Determining how proline oxidase controls its own expression may help us understand how other membrane-bound regulatory proteins work. In addition, the regulatory properties of putA mutations provide a genetic selection for membrane interaction mutants. 2. Flavin dehydrogenase-membrane interactions. Flavin dehydrogenases must associate with the membrane in order to interact properly with the electron transport chain. However, it is not known how they interact with the membrane or what components of the electron transport chain they interact with directly. Genetic studies on the interaction of proline oxidase with the membrane provides a model system for understanding how proteins interact with membranes in vivo. The protein- protein and protein-lipid interactions required for membrane association can be determined by isolation of mutants that prevent proline oxidase from binding to the membrane. Genetic and biochemical analysis of these mutants will indicate what membrane component(s) directly interact with flavin dehydrogenases and what kinds of interactions are required between flavin dehydrogenases and the membrane electron transport chain. Such mutants should provide insight into the general biological problem of how peripheral membrane proteins interact with the membrane.