CAREER: Spectroelectrochemical Studies of the Novel PutA Flavoprotein and its Macromolecular Associations

职业：新型 PutA 黄素蛋白及其大分子关联的光谱电​​化学研究

• 批准号: 0340912
• 负责人: Donald Becker
• 金额: $ 30.9万
• 依托单位: University of Nebraska-Lincoln
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0340912&HistoricalAwards=false
• 关键词: CAREER; Spectroelectrochemical; Studies; Novel; PutA

0091664BeckerThe intriguing flavoprotein, PutA, performs two mutually exclusive functions in Escherichia coli. First, in the cytoplasm, PutA autorepresses transcription of the putA and putP (a proline transporter) genes of the proline utilization (put) regulon. Secondly, PutA associates peripherally with the cytoplasmic membrane where it performs the two-step oxidation of proline to glutamate. The overall goal of this research is to define the mechanism by which PutA switches from a transcriptional regulator to a membrane-associated proline dehydrogenase. The working hypothesis is that the flavin redox state governs PutA macromolecular associations and thus the intracellular location and function of PutA. The objective of this project is to establish the relationship between the flavin redox state and the macromolecular associations of PutA. The first two aims are to investigate the influence of the flavin redox state on the transcriptional regulatory function of PutA and PutA-membrane interactions. The final aim is to correlate conformational changes in PutA with flavin reduction and establish primary structure-function relationships. Experimental approaches will combine spectroelectrochemistry and biophysical methods to provide unique and critical insights into the regulation of PutA. This project will clearly define the influence of the flavin redox state on PutA functions and contribute pivotal understanding into the regulatory mechanism of the multifunctional PutA protein. Studies of proteins that have multiple functions such as PutA are important because they will provide knowledge of a how a particular structural scaffold is used for different purposes. The overall educational goal is to establish an interdisciplinary biochemistry degree program at the University of Missouri-St. Louis. The first step toward this goal is to augment biochemistry curriculum with advanced courses and new laboratory techniques and to broaden research opportunities for undergraduate and graduate students. The study of PutA will promote incorporation of research discoveries and applications into the curriculum.

0091664BeckerThe有趣的黄蛋白Puta在大肠杆菌中执行两个互斥的功能。 首先，在细胞质中，puta自动涂层PUTA和PUTP（PUTP（PUTPON转运蛋白）基因）的转录（PUT）调节。 其次，Puta将其与细胞质膜外周相连，在该膜上将脯氨酸的两步氧化与谷氨酸进行。 这项研究的总体目标是定义Puta从转录调节剂转换为膜相关的脯氨酸脱氢酶的机制。 工作假设是黄素氧化还原状态控制了Puta大分子关联，从而控制了Puta的细胞内位置和功能。 该项目的目的是建立黄素氧化还原国家与Puta的大分子关联之间的关系。 前两个目的是研究黄素氧化还原状态对Puta和Puta-Mmbrane相互作用的转录调节功能的影响。 最终目的是将PUTA中的构象变化与减少黄素相关联并建立主要的结构 - 功能关系。 实验方法将结合光谱化学和生物物理方法，为PUTA调节提供独特而关键的见解。 该项目将清楚地定义黄素氧化还原态对PUTA功能的影响，并为多功能PUTA蛋白的调节机制提供关键的理解。 对具有多种功能（PUTA）的蛋白质的研究很重要，因为它们将提供有关如何将特定结构支架用于不同目的的知识。 总体教育目标是在密苏里街建立跨学科的生物化学学位课程。路易。 朝着这一目标迈进的第一步是通过高级课程和新的实验室技术来扩大生物化学课程，并扩大本科生和研究生的研究机会。 PUTA的研究将促进将研究发现和应用纳入课程中。