The double-histidine Cu²⁺-binding motif: a highly rigid, site-specific spin probe for electron spin resonance distance measurements.
The double-histidine Cu²⁺-binding motif: a highly rigid, site-specific spin probe for electron spin resonance distance measurements.
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DOI：
10.1002/anie.201501968
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发表时间：
2015-05-18
影响因子：
16.6
通讯作者：
Saxena, Sunil
中科院分区：
文献类型：
--
作者：
Cunningham, Timothy F.;Putterman, Miriam R.;Desai, Astha;Horne, W. Seth;Saxena, Sunil
测量远距离距离分布的ESR方法的发展促进了生物物理学的研究。然而，通常使用的自旋标签是高度灵活的，这导致在将ESR测量与蛋白质骨架结构相关联时产生歧义。在这里，我们提出了双组氨酸（dHis） Cu2+结合基序作为双电子-电子共振（DEER）距离测量的刚性自旋探针。该自旋标签由天然氨基酸残基和金属盐原位组装而成，不需要表达后的合成修饰，并且提供的距离分布比常用的蛋白质自旋标签窄得多。基于未标记蛋白质的x射线晶体结构的简单分子建模导致预测的最可能距离在实验值的0.5以内。具有dHis基序的Cu2+ DEER在精确、明确地解决与蛋白质骨架结构和灵活性直接相关的距离限制方面显示出巨大的希望。
The development of ESR methods that measure long-range distance distributions has advanced biophysical research. However, the spin labels commonly employed are highly flexible, which leads to ambiguity in relating ESR measurements to protein-backbone structure. Herein we present the double-histidine (dHis) Cu2+-binding motif as a rigid spin probe for double electron–electron resonance (DEER) distance measurements. The spin label is assembled in situ from natural amino acid residues and a metal salt, requires no postexpression synthetic modification, and provides distance distributions that are dramatically narrower than those found with the commonly used protein spin label. Simple molecular modeling based on an X-ray crystal structure of an unlabeled protein led to a predicted most probable distance within 0.5 of the experimental value. Cu2+ DEER with the dHis motif shows great promise for the resolution of precise, unambiguous distance constraints that relate directly to protein-backbone structure and flexibility.
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DOI：
10.1107/s160053680702288x
发表时间：
2007-06-01
影响因子：
0.9
作者：
Brandi-Blanco, Maria Pilar;de Benavides-Gimenez, Maria Magdalena;Choquesillo-Lazarte, Duane
通讯作者：
Choquesillo-Lazarte, Duane
影响因子：
2.9
作者：
Cunningham, Timothy F.;McGoff, Marshall S.;Saxena, Sunil
通讯作者：
Saxena, Sunil
影响因子：
2
作者：
Finiguerra, Michela G.;Prudencio, Miguel;Huber, Martina
通讯作者：
Huber, Martina
影响因子：
3.2
作者：
Hernandez-Guzman, Jessica;Sun, Li;Warncke, Kurt
通讯作者：
Warncke, Kurt
影响因子：
1
作者：
Hagelueken, Gregor;Ward, Richard;Naismith, James H.;Schiemann, Olav
通讯作者：
Schiemann, Olav