Confining Metal Complexes within Protein Hosts: Models for Metalloprotein Active Sites

这项研究将开发对金属蛋白中的活性部位进行建模的方法，目的是 确定蛋白质如何激活氧气的基本结构-功能关系 严重影响人类健康和老龄化的过程。将制备人造金属蛋白 利用生物素-链霉亲和素技术作为工具，确保特定和可重复的放置 蛋白质宿主中合成的金属络合物。这种方法被认为是一种有效的 模拟天然金属蛋白中活性中心关键性质的方法，包括位点 物种的隔离，初级配位圈的调节，以及对 金属络合物周围的微环境。许多仿生生物的一个明显弱点 这些系统对金属中心周围的微环境的调节能力有限。 没有一个化学系统是在不与当地环境相互作用的情况下孤立运行的。的确有 来自结构生物学的越来越多的证据表明，周围的微环境、空间 金属络合物组成的二次配位球，对 蛋白质的功能，从调节物理性质到递送反应物和 移走产品。 我们的论点是，加强对微环境的监管将导致更好的 了解蛋白质的功能。它还进一步认为，从 本申请中提出的基本分析远远超出了 分子水平的选择性/效率-它们对所有类型的 平台，提供开发仍然缺少的必要信息 功能强大的系统。 我们提出了一种制备人造金属蛋白的方法，该方法允许限制 蛋白质宿主内的合成复合体来调节初级和次级 固定化金属中心周围的配位球。监管这些方面的能力 蛋白质中的配位球将产生系统的结构-功能关系 这将导致对化学过程的更好理解，这些过程与 人类健康。

This research will develop methods to model active sites in metalloproteins for the purpose of determining fundamental structure-function relationships for how proteins activate dioxygen, a process that strongly impacts human health and aging. Artificial metallproteins will be prepared utilizing biotin-streptavidin technology as a tool to ensure specific and reproducible placement of synthetic metal complexes within protein hosts. This approach is proposed to be an effective method to model key properties of the active sites in native metalloproteins, including site isolation of species, regulation of the primary coordination sphere, and control of the microenvironments around the metal complexes. One glaring weakness of many biomimetic systems is their limited ability to regulate the microenvironments that surround metal centers. No chemical system operates in isolation without interacting with its local environment. There is a growing body of evidence from structural biology that the microenvironment, a space around metal complexes that comprises the secondary coordination sphere, has profound effects on protein function that ranges from modulation of physical properties to delivery of reactants and removal of products. It is our contention that the greater regulation of microenvironments will lead to better understanding of protein function. It is further maintained that the benefits gained from fundamental analyses as proposed in this application extend well beyond improvements in selectivities/efficiencies at the molecular level – they are transformative for all types of platforms, providing the requisite information that is still missing for the development of highly functional systems. We propose an approach for preparing artificial metalloproteins that allows for the confinement of synthetic complexes within protein hosts to regulate both the primary and secondary coordination spheres about the immobilized metal centers. The ability to regulate these coordination spheres within a protein will produce systematic structure-function relationships that will lead to an improved understanding of chemical processes that are directly linked to human health.