Stable and rigid DTPA-like paramagnetic tags suitable for in vitro and in situ protein NMR analysis.
Stable and rigid DTPA-like paramagnetic tags suitable for in vitro and in situ protein NMR analysis.
复制标题
稳定且刚性的类 DTPA 顺磁标签，适用于体外和原位蛋白质 NMR 分析。
DOI：
10.1007/s10858-017-0160-3
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发表时间：
2018
影响因子：
2.7
通讯作者：
Su Xun-Cheng
中科院分区：
文献类型：
--
作者：
Chen Jia-Liang;Zhao Yu;Gong Yan-Jun;Pan Bin-Bin;Wang Xiao;Su Xun-Cheng
有机合成对顺磁镧系离子具有高结合亲和力的配体是对蛋白质产生顺磁效应的有效方法。这些在高分辨率核磁共振波谱中表现出的顺磁效应是蛋白质和蛋白质-配体复合物的有价值的动态和结构限制。顺磁标签通常包含金属螯合部分和用于蛋白质修饰的反应基团。在此，我们报告了两种新的类似 DTPA 的标签，4PS-PyDTTA 和 4PS-6M-PyDTTA，它们可以通过稳定的硫醚键位点特异性地附着到蛋白质上。两种蛋白质标签加合物形成稳定的镧系元素复合物，其结合亲和力和顺磁张量相对于吡啶中的 6-甲基基团是可调的。与伪接触位移 (PCS) 相比，评估了 Gd(III) 复合物对蛋白质标签加合物的顺磁弛豫增强 (PRE) 效应，结果表明 4PS-PyDTTA 和 4PS-6M-PyDTTA 标签都是刚性的，并且呈现高质量的 PRE，这对于阐明蛋白质和蛋白质-配体复合物的动力学和相互作用至关重要。我们还表明这两个标签适用于原位蛋白质 NMR 分析。
Organic synthesis of a ligand with high binding affinities for paramagnetic lanthanide ions is an effective way of generating paramagnetic effects on proteins. These paramagnetic effects manifested in high-resolution NMR spectroscopy are valuable dynamic and structural restraints of proteins and protein–ligand complexes. A paramagnetic tag generally contains a metal chelating moiety and a reactive group for protein modification. Herein we report two new DTPA-like tags, 4PS-PyDTTA and 4PS-6M-PyDTTA that can be site-specifically attached to a protein with a stable thioether bond. Both protein-tag adducts form stable lanthanide complexes, of which the binding affinities and paramagnetic tensors are tunable with respect to the 6-methyl group in pyridine. Paramagnetic relaxation enhancement (PRE) effects of Gd(III) complex on protein-tag adducts were evaluated in comparison with pseudocontact shift (PCS), and the results indicated that both 4PS-PyDTTA and 4PS-6M-PyDTTA tags are rigid and present high-quality PREs that are crucially important in elucidation of the dynamics and interactions of proteins and protein-ligand complexes. We also show that these two tags are suitable for in-situ protein NMR analysis.