A Gadolinium Spin Label with Both a Narrow Central Transition and Short Tether for Use in Double Electron Electron Resonance Distance Measurements
A Gadolinium Spin Label with Both a Narrow Central Transition and Short Tether for Use in Double Electron Electron Resonance Distance Measurements
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DOI：
10.1021/acs.inorgchem.8b02892
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发表时间：
2019-03-04
影响因子：
4.6
通讯作者：
Lovett，Janet E.
中科院分区：
文献类型：
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作者：
Shah，Anokhi;Roux，Amandine;Lovett，Janet E.
九坐标含钆自旋标记物[Gd]的设计、合成和应用。介绍了用于EPR光谱纳米距离测量实验的sTPATCN]-SL。自旋标签通过短硫醚系链连接到半胱氨酸，并具有窄的中心跃迁，表明小的零场分裂（ZFS）。一种蛋白同型二聚体TRIM25cc被选择性地用[Gd]标记。sTPATCN]-SL（70%）和一种氮氧化物（30%）在温和条件下，使用商用q波段和自制w波段光谱仪的双电子电子共振（DEER）技术进行测量。该标签通过其有利的弛豫参数和大的Q和w波段的gd （III）间DEER测量的DEER调制深度（9%）显示出极大的希望，可以提高DEER测量的灵敏度，这是在这些条件下记录的最大的。
The design, synthesis, and application of a nine-coordinate gadolinium(III)-containing spin label, [Gd.sTPATCN]-SL, for use in nanometer-distance measurement experiments by EPR spectroscopy is presented. The spin label links to cysteines via a short thioether tether and has a narrow central transition indicative of small zero-field splitting (ZFS). A protein homodimer, TRIM25cc, was selectively labeled with [Gd.sTPATCN]-SL (70%) and a nitroxide (30%) under mild conditions and measured using the double electron electron resonance (DEER) technique with both commercial Q-band and home-built W-band spectrometers. The label shows great promise for increasing the sensitivity of DEER measurements through both its favorable relaxation parameters and the large DEER modulation depth at both Q- and W-band for the inter-Gd(III) DEER measurement which, at 9%, is the largest recorded under these conditions.