RUI: Investigating Salt Induced Effects on RecA Unfolding and Nucleotide Binding
RUI:研究盐对 RecA 解折叠和核苷酸结合的影响
基本信息
- 批准号:0814716
- 负责人:
- 金额:$ 27.55万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Standard Grant
- 财政年份:2008
- 资助国家:美国
- 起止时间:2008-09-01 至 2014-08-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The objective of this project is to identify protein structures that are involved in nucleotide binding and ion protein interactions that result in the high salt induced changes in RecA activity and stability. The Escherichia coli RecA protein is a DNA-dependent ATPase that requires formation of a nucleoprotein filament to perform the DNA strand-exchange reaction in DNA repair and homologous genetic recombination. Salt induced effects that modulate enzyme structure and function involve specific changes in protein structures that result in modified activity and protein-protein interactions. In order to investigate the origin of salt induced effects on RecA the PI will use difference infrared spectroscopy, circular dichroism and fluorescence to study how salt influences the aggregation and unfolding of RecA while simultaneously studying how salt modulates nucleotide binding and ATP hydrolysis. Difference infrared spectroscopy is a powerful yet underutilized technique that has the ability to elucidate changes induced by nucleotide-binding under a variety of solution conditions at the single amino acid level. Combining difference infrared and circular dichroism studies will enable undergraduate students to obtain complementary information about salt induced effects on stability and nucleotide binding. Despite decades of study on many seemingly well-characterized proteins, identifying the specific amino acids and protein structures that are involved in substrate binding, allosteric regulation and ion-mediated processes has continued to be elusive.All of the work in this project will be performed by undergraduates at James Madison University. In addition, the PI has an established history of working with deaf and hard-of-hearing students who will be recruited for research efforts. This project will be important in helping all the involved students gain valuable research experience and will aid in recruiting and retaining future scientists. Furthermore, the results of these studies will lead to a better understanding of not only RecA, but also the wide variety of proteins whose activity, stability and structure are modulated by the presence of salt.
本项目的目的是鉴定参与核苷酸结合和离子蛋白相互作用的蛋白质结构,这些结构导致高盐诱导的RecA活性和稳定性的变化。 大肠杆菌RecA蛋白是DNA依赖性ATP酶,其需要形成核蛋白丝以在DNA修复和同源遗传重组中进行DNA链交换反应。盐诱导的调节酶结构和功能的作用涉及蛋白质结构的特定变化,导致活性改变和蛋白质-蛋白质相互作用。 为了研究盐对RecA的诱导作用的起源,PI将使用差分红外光谱、圆二色谱和荧光来研究盐如何影响RecA的聚集和展开,同时研究盐如何调节核苷酸结合和ATP水解。 差示红外光谱是一种功能强大但未充分利用的技术,能够在单个氨基酸水平上阐明各种溶液条件下核苷酸结合引起的变化。 结合不同的红外和圆二色性研究将使本科生获得盐诱导的稳定性和核苷酸结合的影响的补充信息。 尽管几十年来对许多表面上特征良好的蛋白质进行了研究,但确定参与底物结合,变构调节和离子介导过程的特定氨基酸和蛋白质结构仍然是难以捉摸的。该项目的所有工作将由詹姆斯麦迪逊大学的本科生完成。 此外,PI有一个既定的历史与聋人和听力困难的学生谁将被招募的研究工作。 该项目将有助于帮助所有参与的学生获得宝贵的研究经验,并有助于招募和留住未来的科学家。 此外,这些研究的结果将导致更好地了解不仅RecA,而且各种各样的蛋白质,其活性,稳定性和结构是由盐的存在调节。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Gina MacDonald其他文献
Using Difference Infrared Spectroscopy to Investigate the Effects of pH on PGK-Substrate Complexes
- DOI:
10.1016/j.bpj.2008.12.1560 - 发表时间:
2009-02-01 - 期刊:
- 影响因子:
- 作者:
Adam E. Colbert;Gina MacDonald - 通讯作者:
Gina MacDonald
Concentration and Ion Induced Effects on Nucleotide Binding, Aggregation and Thermal Unfolding Transitions of Reca
- DOI:
10.1016/j.bpj.2009.12.171 - 发表时间:
2010-01-01 - 期刊:
- 影响因子:
- 作者:
Nate D. Talley;Brittany A. Danzig;William R. Cannon;Jennifer S. Martinez;Andrew P. Shreve;Gina MacDonald - 通讯作者:
Gina MacDonald
Using Difference Infrared Spectroscopy to Investigate the Effects of pH on PGK-Substrate Complexes
- DOI:
10.1016/j.bpj.2009.12.4076 - 发表时间:
2010-01-01 - 期刊:
- 影响因子:
- 作者:
Adam E. Colbert;Gina MacDonald - 通讯作者:
Gina MacDonald
The Effect of High Concentration Salt on the Structure, Stability, and Aggregation of RecA
- DOI:
10.1016/j.bpj.2008.12.324 - 发表时间:
2009-02-01 - 期刊:
- 影响因子:
- 作者:
William R. Cannon;Gina MacDonald - 通讯作者:
Gina MacDonald
Gina MacDonald的其他文献
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{{ truncateString('Gina MacDonald', 18)}}的其他基金
Acquisition of a Chiro-optical Spectrophotometric System for Chemistry Research and Student Training
采购用于化学研究和学生培训的手性光学分光光度系统
- 批准号:
0420877 - 财政年份:2004
- 资助金额:
$ 27.55万 - 项目类别:
Standard Grant
PECASE: A Difference Infrared Spectroscopic Study of a Nucleotide Binding Protein
PECASE:核苷酸结合蛋白的差异红外光谱研究
- 批准号:
9733566 - 财政年份:1998
- 资助金额:
$ 27.55万 - 项目类别:
Continuing Grant
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