Cysteine-specific Cu2+ chelating tags used as paramagnetic probes in double electron electron resonance.

Cysteine-specific Cu2+ chelating tags used as paramagnetic probes in double electron electron resonance.
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在双电子电子共振中用作顺磁探针的半胱氨酸特异性 Cu2+ 螯合标签。

DOI:
10.1021/jp5103143
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发表时间:
2015-02-19
期刊:
The journal of physical chemistry. B
影响因子:
--
通讯作者:
Saxena S
Saxena S
中科院分区:
其他
文献类型:
--
作者:
Cunningham TF;Shannon MD;Putterman MR;Arachchige RJ;Sengupta I;Gao M;Jaroniec CP;Saxena S

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双电子电子共振(DEER)是一种有吸引力的技术,用于通过纳米尺度的距离测量来深入了解蛋白质的结构和动力学。在这些测量中最常用的顺磁标记是氮氧自旋标记R1。在这里,我们提出了应用程序的两种类型的高亲和力的Cu 2+螯合标签,基于EDTA和cyclen金属结合基序作为替代的X带DEER探针,使用B1免疫球蛋白结合结构域的蛋白G(GB 1)作为模型系统。这两种类型的标签已被纳入到各种蛋白质二级结构环境,并表现出高光谱灵敏度。特别是,基于环戊二烯基的标签显示的距离分布与同源R1分布相比,具有相当的分布宽度和1-3 μ m内的最可能距离。结果显示cyclen标签作为R1侧链的替代物用于蛋白质中的X带DEER距离测量的可行性。
Double electron electron resonance (DEER) is an attractive technique that is utilized for gaining insight into protein structure and dynamics via nanometer-scale distance measurements. The most commonly used paramagnetic tag in these measurements is a nitroxide spin label, R1. Here, we present the application of two types of high-affinity Cu2+ chelating tags, based on the EDTA and cyclen metal-binding motifs as alternative X-band DEER probes, using the B1 immunoglobulin-binding domain of protein G (GB1) as a model system. Both types of tags have been incorporated into a variety of protein secondary structure environments and exhibit high spectral sensitivity. In particular, the cyclen-based tag displays distance distributions with comparable distribution widths and most probable distances within 1–3 Å when compared to homologous R1 distributions. The results display the viability of the cyclen tag as an alternative to the R1 side chain for X-band DEER distance measurements in proteins.
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