Characterization and evaluation of photolabile (µ-peroxo)(µ-hydroxo)bis[bis(bipyridyl)cobalt caged oxygen compounds to facilitate time-resolved crystallographic studies of cytochrome c oxidase
Emil Sandelin, Jonatan Johannesson, Ola Wendt, Gisela Brändén, Richard Neutze, Carl-Johan Wallentin
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引用次数: 0
Abstract
Photolabile (µ-peroxo)(µ-hydroxo)bis[bis(bipyridyl)-cobalt-based caged oxygen compounds have been synthesized and characterized by optical absorbance spectroscopy, X-ray crystallography. and the quantum yield and redox stability were investigated. Furthermore, conditions were established where redox incompatibilities encountered between caged oxygen compounds and oxygen-dependant cytochrome c oxidase (CcO) could be circumvented. Herein, we demonstrate that millimolar concentrations of molecular oxygen can be released from a caged oxygen compound with spatio-temporal control upon laser excitation, triggering enzymatic turnover in cytochrome c oxidase. Spectroscopic evidence confirms the attainment of a homogeneous reaction initiation at concentrations and conditions relevant for further crystallography studies. This was demonstrated by the oxidizing microcrystals of reduced CcO by liberation of millimolar concentrations of molecular oxygen from a caged oxygen compound. We believe this will expand the scope of available techniques for the detailed investigation of oxygen-dependant enzymes with its native substrate and facilitate further time-resolved X-ray based studies such as wide/small angle X-ray scattering and serial femtosecond crystallography.
促进细胞色素 c 氧化酶时间分辨晶体学研究的可光敏(µ-过氧)(µ-羟基)双[双(联吡啶)钴笼氧化合物的特性和评估
通过光学吸光光谱学和 X 射线晶体学,合成并表征了可光吸收(µ-过氧)(µ-羟基)双[双(联吡啶)-钴基笼式氧化合物,并对其量子产率和氧化还原稳定性进行了研究。此外,我们还确定了可以避免笼氧化合物与依赖氧的细胞色素 c 氧化酶(CcO)之间氧化还原不相容的条件。在这里,我们证明了毫摩尔浓度的分子氧可以在激光激发下从笼氧化合物中释放出来,并具有时空控制能力,从而引发细胞色素 c 氧化酶中的酶转换。光谱学证据证实,在与进一步晶体学研究相关的浓度和条件下,可以实现均匀的反应启动。通过从笼式氧化合物中释放毫摩尔浓度的分子氧来氧化还原型 CcO 的微晶,证明了这一点。我们相信,这将扩大现有技术的范围,以详细研究依赖氧的酶及其原生底物,并促进基于时间分辨 X 射线的进一步研究,例如宽/小角 X 射线散射和串行飞秒晶体学。