Monitoring Complex Formation by Relaxation-Induced Pulse Electron Paramagnetic Resonance Distance Measurements.

Monitoring Complex Formation by Relaxation-Induced Pulse Electron Paramagnetic Resonance Distance Measurements.
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DOI:
10.1002/cphc.201700666
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发表时间:
2017-09-06
期刊:
Chemphyschem : a European journal of chemical physics and physical chemistry
影响因子:
--
通讯作者:
Bode BE
Bode BE
中科院分区:
其他
文献类型:
--
作者:
Giannoulis A;Oranges M;Bode BE

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生物分子复合物通常是多聚体,这刺激了对允许解开其组成和几何排列的方法的需求。脉冲电子顺磁共振(EPR)光谱越来越多地应用于纳米尺度上的几何信息检索。新兴的RIDME(弛豫诱导偶极调制增强)技术在涉及金属中心的距离实验中提供了更高的灵敏度(例如金属蛋白或金属离子标记的蛋白质)。在这里,自旋标记配体与增加量的顺磁性CuII离子的混合物允许在形成的模型复合物中精确定量配体-金属结合。距离测量是高度准确的,并且可以鉴定用于鉴定多聚化的关键方面。除了高精度距离测量之外,量化结合的潜力将进一步扩大EPR的应用范围。
Biomolecular complexes are often multimers fueling the demand for methods that allow unraveling their composition and geometric arrangement. Pulse electron paramagnetic resonance (EPR) spectroscopy is increasingly applied for retrieving geometric information on the nanometer scale. The emerging RIDME (relaxation‐induced dipolar modulation enhancement) technique offers improved sensitivity in distance experiments involving metal centers (e.g. on metalloproteins or proteins labelled with metal ions). Here, a mixture of a spin labelled ligand with increasing amounts of paramagnetic CuII ions allowed accurate quantification of ligand‐metal binding in the model complex formed. The distance measurement was highly accurate and critical aspects for identifying multimerization could be identified. The potential to quantify binding in addition to the high‐precision distance measurement will further increase the scope of EPR applications.
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