Ten Good Reasons for the Use of the Tellurium-Centered Anderson-Evans Polyoxotungstate in Protein Crystallography.
Ten Good Reasons for the Use of the Tellurium-Centered Anderson-Evans Polyoxotungstate in Protein Crystallography.
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DOI：
10.1021/acs.accounts.7b00109
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发表时间：
2017-06-20
影响因子：
18.3
通讯作者：
Rompel A
中科院分区：
文献类型：
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作者：
Bijelic A;Rompel A
蛋白质晶体学是目前在原子分辨率范围内获得靶蛋白和蛋白质配体复合物结构信息的最有效和最广泛使用的方法。以这种方式获得的知识对于理解蛋白质的生物学、化学和生物化学及其功能至关重要，而且对于开发具有高度药理和医学意义的化合物也至关重要。在这里，我们解决了蛋白质晶体学的核心问题：结晶过程的不可预测性。获得高分辨率的蛋白质晶体是通过x射线晶体学进行任何结构研究的必要步骤；然而，这种方法仍然基本上依赖于试验和错误，使其成为一个非常耗时和消耗资源的过程。为了获得高质量的晶体，添加剂的使用是一种既定的过程，以使或改善蛋白质的结晶。因此，需要一种更通用的添加剂来处理更广泛的蛋白质，因为它将代表蛋白质晶体学的巨大进步，同时极大地影响多个研究领域。这反过来又可以为整个社会带来整体利益，因为它受益于新药物或改良药物的更快发展，以及对生物、生化和药理学现象的更深入理解。为此，我们测试了几种属于新兴多金属氧酸盐（pom）的化合物作为结晶添加剂的适用性，并发现以碲为中心的Anderson-Evans多钨氧酸盐[TeW6O24]6 - (TEW)是最合适的pom原型。在TEW作为结晶添加剂的首次成功应用后，我们反复报道了TEW对蛋白质结晶行为的积极影响，并特别关注了蛋白质- TEW的相互作用。由于静电相互作用是TEW与蛋白质结合的主要力量，因此具有高度负电荷的TEW原则上可以处理所有具有正电荷贴片的蛋白质。此外，由于其高度的结构和化学多样性，TEW与一些常用的结晶添加剂相比具有很大的优势。因此，我们总结了TEW有利于蛋白质结晶的所有特点，并提出了TEW作为一种强大的添加剂在蛋白质结晶学中推广使用的十大理由。我们的结果表明，TEW是一种化合物，在许多方面，是注定的结晶添加剂。我们认为许多晶体学家，特别是研究人员，他们不是该领域的专家，但愿意结晶他们结构未知的目标蛋白，可以从使用TEW中受益，因为它能够通过提供有价值的异常信号来促进结晶过程本身和随后的结构解析，这有助于分相步骤。
Protein crystallography represents at present the most productive and most widely used method to obtain structural information on target proteins and protein–ligand complexes within the atomic resolution range. The knowledge obtained in this way is essential for understanding the biology, chemistry, and biochemistry of proteins and their functions but also for the development of compounds of high pharmacological and medicinal interest. Here, we address the very central problem in protein crystallography: the unpredictability of the crystallization process. Obtaining protein crystals that diffract to high resolutions represents the essential step to perform any structural study by X-ray crystallography; however, this method still depends basically on trial and error making it a very time- and resource-consuming process. The use of additives is an established process to enable or improve the crystallization of proteins in order to obtain high quality crystals. Therefore, a more universal additive addressing a wider range of proteins is desirable as it would represent a huge advance in protein crystallography and at the same time drastically impact multiple research fields. This in turn could add an overall benefit for the entire society as it profits from the faster development of novel or improved drugs and from a deeper understanding of biological, biochemical, and pharmacological phenomena. With this aim in view, we have tested several compounds belonging to the emerging class of polyoxometalates (POMs) for their suitability as crystallization additives and revealed that the tellurium-centered Anderson–Evans polyoxotungstate [TeW6O24]6– (TEW) was the most suitable POM-archetype. After its first successful application as a crystallization additive, we repeatedly reported on TEW’s positive effects on the crystallization behavior of proteins with a particular focus on the protein–TEW interactions. As electrostatic interactions are the main force for TEW binding to proteins, TEW with its highly negative charge addresses in principle all proteins possessing positively charged patches. Furthermore, due to its high structural and chemical diversity, TEW exhibits major advantages over some commonly used crystallization additives. Therefore, we summarized all features of TEW, which are beneficial for protein crystallization, and present ten good reasons to promote the use of TEW in protein crystallography as a powerful additive. Our results demonstrate that TEW is a compound that is, in many respects, predestined as a crystallization additive. We assume that many crystallographers and especially researchers, who are not experts in this field but willing to crystallize their structurally unknown target protein, could benefit from the use of TEW as it is able to promote both the crystallization process itself and the subsequent structure elucidation by providing valuable anomalous signals, which are helpful for the phasing step.