Molecular insights into evolution and mechanisms of binding, release and electron transfer between plastocyanin or cytochrome c6 and photosystem I

对质体蓝素或细胞色素 c6 与光系统 I 之间结合、释放和电子转移的进化和机制的分子见解

• 批准号: 37189714
• 负责人: Professor Dr. Michael Hippler
• 金额: --
• 依托单位: Arbeitsgruppe Plant Biochemistry and Biotechnology
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2011-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/37189714?language=en
• 关键词: Molecular; insights; into; evolution; mechanisms

The electron transfer reaction between plastocyanin (pc) or cytchrome c6 (cyt c6) and photosystem I (PSI) in green algae provides a unique system to gain molecular insights in general principles of protein-protein recognition. It is an attractive molecular model to investigate how electron transfer is optimized between partners in vitro and under physiological in vivo conditions where concentration of reaction partners, the pH value, the ionic strength of the solution and other parameter underlie permanent changes. We will perform site-directed mutagenesis of the plastid-encoded genes for PSI subunits PsaA/PsaB and nuclear encoded cyt c6 as well as plastid and nuclear transformation and heterologous expression of cyt c6 to alter specific residues that are important for optimal binding/ release and electron transfer between the donors and PSI. The electron transfer reaction will be analysed in vitro and in vivo by fast absorbance spectroscopy as well as chemical crosslinking techniques and mass spectrometry. It is our goal to provide indepth mechanistic understanding of how (i) binding, release and electron transfer were optimized during evolution, (ii) to provide a detailed molecular model and (iii) understand how these optimizations are reflected in the physiological performance of the alga.

绿藻中叶绿体青素（pc）或细胞色素c6 （cyt c6）与光系统I （PSI）之间的电子转移反应提供了一个独特的系统来获得蛋白质-蛋白质识别的一般原理的分子见解。这是一个有吸引力的分子模型，用于研究在体外和体内生理条件下，在反应伙伴的浓度、pH值、溶液的离子强度和其他参数构成永久变化的基础上，电子转移是如何优化的。我们将对PSI亚基PsaA/PsaB和核编码cyt c6的质体编码基因进行定点突变，以及质体和核转化和cyt c6的异源表达，以改变对供体和PSI之间的最佳结合/释放和电子转移至关重要的特定残基。电子转移反应将在体外和体内通过快速吸收光谱以及化学交联技术和质谱分析。我们的目标是深入了解(1)结合、释放和电子转移在进化过程中是如何优化的，(2)提供详细的分子模型，(3)了解这些优化是如何反映在藻类的生理性能上的。