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Mechanisms of Energy Conservation in Bifurcating Electron Transfer Flavoproteins

分叉电子转移黄素蛋白的能量守恒机制

基本信息

批准号：
1808433
负责人：
Anne-Frances Miller
金额：
$ 47.1万
依托单位：
University of Kentucky Research Foundation
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-01 至 2021-12-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1808433&HistoricalAwards=false
关键词：
Mechanisms Energy Conservation Bifurcating Electron

项目摘要

Just as electrical wires carry power to every room in our houses, cells have dedicated proteins carrying a current of electrons from reactions that generate electrons to vital reactions that require them. This project addresses a newly-recognized class of 'electron transferring flavoproteins' (Etfs) that act as energy brokers, trading quantity for quality by accepting pairs of modest-energy electrons and concentrating their energy onto just one of the pair to produce one high-energy electron. Crucially, this biochemical 'step-up station' makes it possible for cells to fix nitrogen gas from air to generate their own fertilizer, making food production possible where it otherwise would not be. The project seeks to understand HOW these Etfs accomplish this complex process. The research seeks to learn what is special about the site at which energy is reallocated among the two electrons by studying the molecule on which this happens (this flavin is a derivative of the riboflavin vitamin B2). The research also seeks to identify safety mechanisms built into the protein scaffold that allow such demanding transformations to occur within a benign, non-toxic protein. Thus the work aims to articulate the underlying principles of the process, so that they can be designed into human-made devices and materials, to increase our ability to use solar power and boost the efficiency and versatility with which we use electrical energy in general. Flavins are just one example of many plant pigments that present powerful chemical properties along with glorious colors. The project scientists share their enthusiasm for pigments via a series of workshops and a course on plant pigments, fibers and fragrances in which participants create a fiber art project using natural materials while learning about the chemical principles underlying plant colors. This creative chemistry course gives all participants a chance to design experiments and integrate science and art.A long-familiar family of proteins has recently been found to have a surprising capacity to re-allocate energy among pairs of electrons acquired from NADH, yielding one with more reducing power than the NADH source (electron transfer bifurcation, or 'bifurcation'). This research establishes which of the two flavins plays which of the contrasting roles identified in the newly-purified bifurcating electron transferring flavoprotein (Etf), and then seeks to understand what aspects of the protein environment are responsible for each of the different activities (conventional electron transfer vs. bifurcation). Spectroelectrochemical titrations measure the extent to which individual amino acids change the flavin reduction potentials (Eo) in proteins bearing amino acid substitutions, and spectroscopic studies elucidate changes in flavin covalent bonding, hydrogen bonding and protonation states, mapping out the active site's role in producing bifurcating activity. Residues to be targeted include a cysteine common to all the Etfs associated with nitrogen fixation but not conserved in others. Thus the research may reveal the protein context needed in order to exploit flavins' capacity for this reactivity while suppressing side-effects. The objective is to elucidate principles underlying the versatility and efficiency of bifurcation, for implementation beyond biochemistry.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

就像电线把电力输送到我们房子的每个房间一样，细胞也有专门的蛋白质，从产生电子的反应到需要电子的生命反应中携带电子流。该项目研究了一种新发现的“电子转移黄素蛋白”（Etfs），它充当能量经纪人，通过接受一对中等能量的电子并将其能量集中在其中一个电子上产生一个高能电子来进行数量交换。至关重要的是，这种生物化学“升压站”使细胞能够从空气中固定氮气来产生自己的肥料，使粮食生产成为可能，否则它不会。该项目旨在了解这些ETF如何完成这一复杂的过程。该研究旨在通过研究发生这种情况的分子（这种黄素是核黄素维生素B2的衍生物）来了解能量在两个电子之间重新分配的位置的特殊之处。该研究还试图确定蛋白质支架中内置的安全机制，这些机制允许在良性无毒蛋白质中发生这种要求苛刻的转化。因此，这项工作旨在阐明该过程的基本原理，以便将其设计成人造设备和材料，以提高我们使用太阳能的能力，并提高我们使用电能的效率和多功能性。黄素只是许多植物色素中的一个例子，它们具有强大的化学特性，沿着着绚丽的色彩。项目科学家通过一系列关于植物色素、纤维和香料的研讨会和课程分享他们对色素的热情，参与者在学习植物颜色背后的化学原理的同时，使用天然材料创建纤维艺术项目。这门创造性的化学课程为所有参与者提供了设计实验和整合科学与艺术的机会。最近发现一个长期熟悉的蛋白质家族具有惊人的能力，可以在从NADH获得的电子对之间重新分配能量，产生一个比NADH源更具有还原能力的电子（电子转移分叉，或“分叉”）。这项研究确定了两种黄素中的哪一种在新纯化的分叉电子转移黄素蛋白（Etf）中起着相反的作用，然后试图了解蛋白质环境的哪些方面负责每个不同的活动（常规电子转移与分叉）。光谱电化学滴定测量在何种程度上个别氨基酸改变黄素还原电位（Eo）在蛋白质轴承氨基酸取代，光谱研究阐明黄素共价键，氢键和质子化状态的变化，绘制出的活性位点的作用，在生产分叉活动。待靶向的残基包括与固氮相关的所有Etf共有的但在其他Etf中不保守的半胱氨酸。因此，这项研究可能揭示了利用黄素的这种反应能力所需的蛋白质背景，同时抑制副作用。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。