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引用次数: 0
摘要
促进磷酸基团转移的酶构成了所有生命王国中最广泛的蛋白质家族。它们约占人类基因组中蛋白质的 10%。了解酶催化这些反应的机制,对于确定它们调控过程的特征至关重要。金属氟化物可用作研究这些酶的多功能工具。这些离子型物质与磷酸盐和转移的磷酸基带有相同的电荷,此外,它们还能使酶处于重要的催化状态,具有光谱敏感性的原子直接与活性位点残基相互作用。这些磷酸盐代用品的离子性质还允许将其移除并用其他类似物替代。在此,我们介绍了获得这些复合物的最佳方法,以及它们在核磁共振、X 射线晶体学、低温电子显微镜和 SAXS 中的应用,并描述了一种新的金属氟化物--四氟化钪,它在使用软 X 射线时具有显著的异常信号。
Metal fluorides—multi-functional tools for the study of phosphoryl transfer enzymes, a practical guide
Enzymes facilitating the transfer of phosphate groups constitute the most extensive protein families across all kingdoms of life. They make up approximately 10% of the proteins found in the human genome. Understanding the mechanisms by which enzymes catalyze these reactions is essential in characterizing the processes they regulate. Metal fluorides can be used as multifunctional tools to study these enzymes. These ionic species bear the same charge as phosphate and the transferring phosphoryl group and, in addition, allow the enzyme to be trapped in catalytically important states with spectroscopically sensitive atoms interacting directly with active site residues. The ionic nature of these phosphate surrogates also allows their removal and replacement with other analogs. Here, we describe the best practices to obtain these complexes, their use in NMR, X-ray crystallography, cryo-EM, and SAXS and describe a new metal fluoride, scandium tetrafluoride, which has significant anomalous signal using soft X-rays.
期刊介绍:
Structure aims to publish papers of exceptional interest in the field of structural biology. The journal strives to be essential reading for structural biologists, as well as biologists and biochemists that are interested in macromolecular structure and function. Structure strongly encourages the submission of manuscripts that present structural and molecular insights into biological function and mechanism. Other reports that address fundamental questions in structural biology, such as structure-based examinations of protein evolution, folding, and/or design, will also be considered. We will consider the application of any method, experimental or computational, at high or low resolution, to conduct structural investigations, as long as the method is appropriate for the biological, functional, and mechanistic question(s) being addressed. Likewise, reports describing single-molecule analysis of biological mechanisms are welcome.
In general, the editors encourage submission of experimental structural studies that are enriched by an analysis of structure-activity relationships and will not consider studies that solely report structural information unless the structure or analysis is of exceptional and broad interest. Studies reporting only homology models, de novo models, or molecular dynamics simulations are also discouraged unless the models are informed by or validated by novel experimental data; rationalization of a large body of existing experimental evidence and making testable predictions based on a model or simulation is often not considered sufficient.