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Design of electron conductive protein-protein interfaces and redox-active multiprotein assemblies

电子传导蛋白质-蛋白质界面和氧化还原活性多蛋白质组装体的设计

基本信息

批准号：
393131496
负责人：
Dr. Julian Esselborn
金额：
--
依托单位：
Department of Chemistry and Biochemistry
依托单位国家：
德国
项目类别：
Research Fellowships
财政年份：
2017
资助国家：
德国
起止时间：
2016-12-31 至 2019-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/393131496?language=en
关键词：
Design electron conductive protein interfaces

项目摘要

This project aims at designing artificial protein interfaces using metals as binding agent between proteins. This should lead to stable protein-protein complexes and at the same time to electron conductivity between the reduction/oxidation sites in the centre of the complexed proteins via the metal at the interface. To allow for reduction/oxidation of the interfacial metals, three different metals in the form of copper ions, iron ions as central atom in heme cofactors and iron/sulfur clusters of the [4Fe4S] type shall be tested, as these are the major agents of metal-based biological electron transfer capability in nature.Each of these conductive elements needs a different and highly specialized protein binding site, which will be designed with computational tools, before the resulting proteins will be produced using Escherichia coli and purified ex vitro. Subsequently the formation of protein-protein complexes and reduction/oxidation activity will be investigated upon addition of the respective metal/metal-cofactor. Once one of the three approaches is successful in generating protein dimers with electron conductivity over the protein-protein interface, the method will be expanded to the self-assembly of larger arrays. For other proteins with non-condcutive metals, the formation of highly ordered arrays of several thousand proteins reaching µm sized sheets by using several protein binding sites in proper orientation towards each other has already been demonstrated.Since electron consuming or electron discharging reactions catalyzed by highly specialized enzymes are of biotechnological interest, the coupling of these reactions to either electrodes or other enzymes, which can consume cheap substrates like hydrogen or even light, is an important goal in biochemistry. However, methods to "wire" proteins together are not well developed and shall thus be established in this project.Larger conductive assemblies of proteins can serve as research tools in the future to understand longer-range electron transfer in nature and might be developed into valuable biological building blocks for electric circuits on a nm to µm scale.

该项目旨在设计人工蛋白质界面，使用金属作为蛋白质之间的结合剂。这将导致稳定的蛋白质-蛋白质复合物，同时通过界面处的金属在复合蛋白质中心的还原/氧化位点之间产生电子传导性。为了允许界面金属的还原/氧化，应测试三种不同的金属，其形式为铜离子、作为血红素辅因子中的中心原子的铁离子和[4Fe 4S]型的铁/硫簇，因为这些是自然界中基于金属的生物电子转移能力的主要因素。这些导电元素中的每一种都需要不同且高度专业化的蛋白质结合位点，其将用计算工具设计，然后使用大肠杆菌生产所得蛋白质并体外纯化。随后，蛋白质-蛋白质复合物的形成和还原/氧化活性将在添加相应的金属/金属辅因子后进行研究。一旦这三种方法中的一种成功地在蛋白质-蛋白质界面上产生具有电子传导性的蛋白质二聚体，该方法将扩展到更大阵列的自组装。对于具有非导电金属的其他蛋白质，已经证明了通过使用彼此适当取向的几个蛋白质结合位点，可以形成数千个蛋白质的高度有序阵列，达到μm大小的片层。由于由高度专门化的酶催化的电子消耗或电子释放反应具有生物技术意义，因此将这些反应与电极或其他酶耦合，它可以消耗像氢甚至光这样的廉价底物，这是生物化学的一个重要目标。然而，将蛋白质“连接”在一起的方法还没有得到很好的发展，因此在本项目中应建立更大的蛋白质导电组装体，将来可以作为研究工具，以了解自然界中更长距离的电子转移，并可能被开发为纳米至微米尺度的电路的有价值的生物构建模块。