NMR STUDIES OF BACTERIAL REGULATORY PROTEINS

细菌调节蛋白的核磁共振研究

• 批准号: 2824632
• 负责人: John Cavanagh
• 金额: $ 20.61万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2824632
• 关键词: Bacillus subtilis; DNA binding protein; bacterial proteins; biological signal transduction; conformation; intermolecular interaction; molecular dynamics; mutant; nuclear magnetic resonance spectroscopy; phosphorylation; protein structure function; site directed mutagenesis; spores; thermodynamics; transcription factor

DESCRIPTION: Research is focused towards understanding the mechanism of action and molecular recognition features of proteins involved in bacterial protection. Energetic and biochemical methods, combined with NMR spectroscopy, are used to investigate the actions and interactions of two classes of proteins: (i) transition state regulators and (ii) response regulators. Transition state regulators are able to control bacterial "decision making" by their ability to recognize and bind to a diverse array of DNA sequences. Response regulators constitute one-half of a ubiquitous communication module known as a two-component switch. Two-component switches are found in all bacteria and have been shown to be vital components in all signal transduction pathways leading to the development of virulence. As a model system, the signal transduction pathway that initiates sporulation in B. subitlis has been chosen. In response to nutrient deprivation and/or high cell density, B. subtilis passes from its vegetative-state, through a transition-state, to a self-protective stationary phase that involves the development of a spore. This system contains the transition-state regulator AbrB and a pair of two-component switches, including the response regulators Spo0F and Spo0A. It is regarded as a paradigm for bacterial signal transduction systems required for virulence, environmental sensing, cell-cycle control and cell-cell communication. The studies are designed to: (a) determine the DNA binding nature of the transition state regulator AbrB and to explain its mechanism of widespread DNA recognition. Initial investigations have revealed a putative novel DNA-binding motif which may have the capacity to adopt different conformations due to the flexible nature of the protein backbone; (b) address critical issues of molecular recognition and mechanisms involving response regulator proteins. Both the molecular recognition properties and many of the mechanistic features of the response regulators Spo0F and Spo0A have recently been shown to depend, not only on their structure, but also equally on their inherent dynamic characteristics. Moreover, the investigations have allowed Dr. Cavanagh to propose practical models for response regulator action. The research described in the proposal is designed to test the veracity of these models, subsequently extend them, and provide general insight into the two-component mode of action.

描述：研究重点是理解 涉及细菌的蛋白质的作用和分子识别特征 保护。 能量和生化方法，结合NMR 光谱法用于研究两个 蛋白质类别：（i）过渡状态调节剂和（ii）响应 监管机构。 过渡状态调节器能够控制细菌 “决策”通过他们识别和结合多样的数组的能力 DNA序列。 响应调节剂构成无处不在的一半 通信模块称为两个组件开关。 两个组件 在所有细菌中都发现开关，并且已被证明至关重要 所有信号转导途径中的组件导致发展 毒力。 作为模型系统，信号转导途径 已经选择了B. subitlis中的孢子形成。 响应 营养剥夺和/或高细胞密度，枯草芽孢杆菌从其 通过过渡状态到自我保护的营养状态 涉及孢子发育的固定阶段。 这个系统 包含过渡状态调节器ABR和一对两组 开关，包括响应调节器SPO0F和SPO0A。 它被认为 作为细菌信号转导系统的范式 毒力，环境传感，细胞周期控制和细胞细胞 沟通。 研究的设计为：（a）确定DNA的结合性质 过渡状态调节器ABRB并解释其广泛的机制 DNA识别。 最初的调查显示了一部假定的小说 DNA结合图案可能有能力采用不同 由于蛋白质主链的柔性性质而引起的构象； （b） 解决分子识别和涉及的机制的关键问题 响应调节蛋白。 分子识别特性和 响应调节器SPO0F和SPO0A的许多机械特征 最近已证明不仅取决于其结构，还取决于其结构 同样，它们固有的动态特征。 而且， 调查使Cavanagh博士可以为 响应调节器作用。 提案中描述的研究是 旨在测试这些模型的真实性，随后将其扩展，并 提供对两组分动作方式的一般洞察力。