Fingerprinting redox and ligand states in haemprotein crystal structures using resonance Raman spectroscopy
Fingerprinting redox and ligand states in haemprotein crystal structures using resonance Raman spectroscopy
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DOI：
10.1107/s1399004714004039
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发表时间：
2014-05-01
期刊：
影响因子：
--
通讯作者：
Hough, Michael A.
中科院分区：
文献类型：
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作者：
Kekilli, Demet;Dworkowski, Florian S. N.;Hough, Michael A.
为具有活性氧化还原中心的蛋白质的晶体结构分配正确的氧化还原态和配体态是获得有效的功能信息和防止结构误解的关键。单晶光谱，特别是在大分子晶体学光束线原位应用时，允许在收集结构数据期间对氧化还原和配体状态进行光谱研究，并鉴定蛋白质晶体中的反应中间体。用Alcaligenes xylosoxidans的细胞色素c′在瑞士光源X10SA光束线上进行了单晶共振拉曼光谱和大分子晶体学的结合。这允许指纹识别和验证不同的氧化还原和配体状态，识别振动模式和中间体识别以及监测辐射引起的变化。这种结合的方法提供了一个强大的工具来获得互补数据，并正确分配氧化还原蛋白晶体中的真实氧化和配体状态。
It is crucial to assign the correct redox and ligand states to crystal structures of proteins with an active redox centre to gain valid functional information and prevent the misinterpretation of structures. Single-crystal spectroscopies, particularly when applied in situ at macromolecular crystallography beamlines, allow spectroscopic investigations of redox and ligand states and the identification of reaction intermediates in protein crystals during the collection of structural data. Single-crystal resonance Raman spectroscopy was carried out in combination with macromolecular crystallography on Swiss Light Source beamline X10SA using cytochrome c' from Alcaligenes xylosoxidans. This allowed the fingerprinting and validation of different redox and ligand states, identification of vibrational modes and identification of intermediates together with monitoring of radiation-induced changes. This combined approach provides a powerful tool to obtain complementary data and correctly assign the true oxidation and ligand state(s) in redox-protein crystals.