MOLECULAR BIOLOGY OF MOT AND THE CHE GENE PRODUCTS

MOT 的分子生物学和 CHE 基因产品

• 批准号: 2390262
• 负责人: PHILIP MATSUMURA
• 金额: $ 29.49万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2390262
• 关键词: X ray crystallography; biological signal transduction; chemoreceptors; chemotaxis; conformation; enzyme activity; enzyme complex; enzyme linked immunosorbent assay; flagellum; membrane proteins; mutant; nuclear magnetic resonance spectroscopy; phosphoprotein phosphatase; phosphorylation; protein structure function; solutions; structural biology; suppressor mutations; temperature sensitive mutant

Two-component signalling is a widespread biological phenomenon found primarily in procaryotes but also in eucaryotes. Its abundant representation makes it clear that virtually all phenomena studied in bacteria are either directly or indirectly regulated by at least one two- component system. Furthermore, since it appears that all members within each component family share structural and function homology, there is a unifying theme which has allowed previously unrelated biological phenomena to share findings. This type of informational networking is rare and has accelerated the progress in many two-component systems. For example, results from seemingly diverse areas as osmoregulation (OmpR), sporulation (SpoOA/F) or chemotaxis (CheY) have been compared and used to gain insight into each particular system. Also, new members in the 100+ family of CheY homologs can immediately benefit from the cumulative database. Clearly, there are differences and variations but the basic motif appears to be retained. This proposal is focused primarily on the structure and function of CheY. CheY is in the fortunate position of being the only response regulator for which there is high resolution structural information to combine with extensive genetic and biochemical data. We have both solid-state crystallographic and solution NMR structures of CheY. In collaboration with Dr. F.W. Dahlquist, we propose to continue our efforts in solving the solution NMR structure of the phosphorylated form of CheY and defining the molecular mechanism of activation. In addition, we will further elucidate the sites of protein-protein interaction and the conversion of kinase binding domain to the activated flagellar motor binding domain of CheY. We will also study the enhancement of the phosphatase activity of CheZ when it is bound to CheAS. These ambitious experiments represent a fortuitous combination of events; the development of a biologically important signal transduction system, an overproduced and well characterized response regulator protein, and the newly emerging technical advances in crystallography and NMR spectroscopy.

两个组件信号传导是一种广泛的生物学现象 主要在丙烯酸酯中，也是桉树。 它丰富 表示明确表明，实际上所有研究的现象 细菌直接或间接地由至少一个两种 组件系统。 此外，由于看来所有成员 每个组件都共享结构和功能同源性，有一个 统一主题允许以前无关的生物学现象 分享发现。 这种信息网络非常罕见，并且 加速了许多两个组件系统中的进度。 例如， 似乎来自渗透压（OMPR），孢子形成的不同地区的结果 （SPOOA/F）或趋化性（Chey）已被比较并用于获得见解 进入每个特定系统。 此外，夏的100+家族的新成员 同源物可以立即受益于累积数据库。 清楚地， 存在差异和变化，但基本主题似乎是 保留。 该提案主要集中在Chey的结构和功能上。 Chey处于幸运的位置，是唯一的响应调节器 有高分辨率的结构信息要与 广泛的遗传和生化数据。 我们都有固态 Chey的晶体学和溶液NMR结构。 合作 与F.W. Dahlquist博士一起，我们建议继续我们的努力来解决 溶液NMR的磷酸化形式的Chey的结构，并定义 激活的分子机制。 此外，我们将进一步阐明 蛋白质 - 蛋白质相互作用的位点和激酶的转化 结合了Chey活化的鞭毛运动结合结构域。 我们还将研究Chez的磷酸酶活性的增强 当它一定会伴侣时。 这些雄心勃勃的实验代表了事件的偶然结合。 生物学上重要的信号转导系统的发展，一个 过度生产和表征良好的响应调节蛋白，以及 晶体学和NMR光谱法新出现的技术进步。