Monitoring and validating active site redox states in protein crystals
Monitoring and validating active site redox states in protein crystals
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DOI：
10.1016/j.bbapap.2010.12.017
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发表时间：
2011-06-01
影响因子：
3.2
通讯作者：
Hough, Michael A.
中科院分区：
文献类型：
--
作者：
Antonyuk, Svetlana V.;Hough, Michael A.
利用同步辐射进行高分辨率蛋白质晶体学研究是现代生物学中最有力的工具之一。分辨率的提高是由于使用了具有更高亮度和通量的X射线束线以及开发了先进的探测器。然而，人们越来越认识到，这些进步带来的好处有相关的成本，即X射线辐射对样品的有害影响（辐射损伤）。特别是，X射线诱导的还原和氧化还原中心的损害已被证明比其他辐射损害效应（如分辨率损失或二硫键损害）发生得更快。在晶体学实验过程中选择适当的原位单晶光谱组合，如紫外可见吸收和X射线吸收光谱（XAFS），可以有效地监测蛋白质晶体中的氧化还原状态，同时进行结构测定。在通过暴露于X射线束产生催化中间体物质的情况下，这种方法也是必不可少的。在这篇文章中，我们提供了一些例子，其中多个单晶光谱是理解晶体结构中Fe和Cu中心氧化还原状态的关键。这篇文章是特刊的一部分，题为：蛋白质结构和功能的晶体状态。(C)2011 Elsevier B.V.保留所有权利。
High resolution protein crystallography using synchrotron radiation is one of the most powerful tools in modern biology. Improvements in resolution have arisen from the use of X-ray beamlines with higher brightness and flux and the development of advanced detectors. However, it is increasingly recognised that the benefits brought by these advances have an associated cost, namely deleterious effects of X-ray radiation on the sample (radiation damage). In particular, X-ray induced reduction and damage to redox centres has been shown to occur much more rapidly than other radiation damage effects, such as loss of resolution or damage to disulphide bridges. Selection of an appropriate combination of in-situ single crystal spectroscopies during crystallographic experiments, such as UV-visible absorption and X-ray absorption spectroscopy (XAFS), allows for effective monitoring of redox states in protein crystals in parallel with structure determination. Such approaches are also essential in cases where catalytic intermediate species are generated by exposure to the X-ray beam. In this article, we provide a number of examples in which multiple single crystal spectroscopies have been key to understanding the redox status of Fe and Cu centres in crystal structures. This article is part of a Special Issue entitled: Protein Structure and Function in the Crystalline State. (C) 2011 Elsevier B.V. All rights reserved.