Photoreduction and validation of haem-ligand intermediate states in protein crystals by in situ single-crystal spectroscopy and diffraction.
Photoreduction and validation of haem-ligand intermediate states in protein crystals by in situ single-crystal spectroscopy and diffraction.
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DOI：
10.1107/s2052252517002159
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发表时间：
2017-05-01
期刊：
影响因子：
3.9
通讯作者：
Hough MA
中科院分区：
文献类型：
--
作者：
Kekilli D;Moreno-Chicano T;Chaplin AK;Horrell S;Dworkowski FSN;Worrall JAR;Strange RW;Hough MA
综合结构生物学可以产生强大的协同作用，并最大限度地获得生物信息。本文介绍了将x射线晶体学与单晶共振拉曼光谱和紫外可见光谱相结合研究血红素蛋白功能的两个例子。结合多种方法研究晶体形式的蛋白质，可以获得强大的协同效应。探测晶体中x射线晶体结构的相同区域的光谱学可以深入了解氧化还原，配体和自旋状态，以补充从电子密度图中获得的信息。正确的晶体结构分配到正确的蛋白质氧化还原和配体状态是必不可少的，以避免误读结构数据。这对于血红素蛋白来说是一个特别值得关注的问题，血红素蛋白可以占据广泛的氧化还原状态，并且对由x射线与晶体中的水分子相互作用产生的溶剂化电子而被还原非常敏感。本文应用单晶光谱指纹技术研究了细胞色素c′中铁血红素的激光光还原。此外，在原位x射线驱动生成血红素中间体的晶体中的染料脱色型过氧化物酶A （DtpA）从lividans描述。
Integrated structural biology can yield powerful synergies and maximize the biological information gained. Two examples are described of combining X-ray crystallography with single-crystal resonance Raman and UV–visible spectroscopies to study the functions of haem proteins. Powerful synergies are available from the combination of multiple methods to study proteins in the crystalline form. Spectroscopies which probe the same region of the crystal from which X-ray crystal structures are determined can give insights into redox, ligand and spin states to complement the information gained from the electron-density maps. The correct assignment of crystal structures to the correct protein redox and ligand states is essential to avoid the misinterpretation of structural data. This is a particular concern for haem proteins, which can occupy a wide range of redox states and are exquisitely sensitive to becoming reduced by solvated electrons generated from interactions of X-rays with water molecules in the crystal. Here, single-crystal spectroscopic fingerprinting has been applied to investigate the laser photoreduction of ferric haem in cytochrome c′. Furthermore, in situ X-ray-driven generation of haem intermediates in crystals of the dye-decolourizing-type peroxidase A (DtpA) from Streptomyces lividans is described.