A Chiral Lanthanide Tag for Stable and Rigid Attachment to Single Cysteine Residues in Proteins for NMR, EPR and Time-Resolved Luminescence Studies.
A Chiral Lanthanide Tag for Stable and Rigid Attachment to Single Cysteine Residues in Proteins for NMR, EPR and Time-Resolved Luminescence Studies.
复制标题
DOI：
10.1002/chem.202101143
复制
发表时间：
2021-09-09
期刊：
影响因子：
--
通讯作者：
Butler SJ
中科院分区：
文献类型：
--
作者：
Herath ID;Breen C;Hewitt SH;Berki TR;Kassir AF;Dodson C;Judd M;Jabar S;Cox N;Otting G;Butler SJ
一个特定于蛋白质的稀土结合标签提供了一种通过多种光谱技术探测蛋白质的工具，包括核磁共振、电子顺磁共振和时间分辨发光光谱。这里提出了一种新的稳定的手性LnIII标记，称为C12，用于与半胱氨酸残基的自发定量反应生成稳定的硫醚键。标签的合成方案相对简单，标签储存和运输都很稳定。它极大地增强了对硒半胱氨酸的反应性，为在含有半胱氨酸残基的蛋白质中选择性标记硒半胱氨酸开辟了一条途径。C12标签负载了TbIII或TmIII离子，很容易在蛋白质核磁共振谱中产生伪接触位移(PCS)。它在稀土元素和蛋白质之间产生了一种相对刚性的链结，这有利于用单磁化率各向异性张量来解释PCSS，并适用于测量双电子共振实验中的距离分布。当与半胱氨酸或其他硫醇化合物反应时，TbIII络合物的发光量子产率提高了100倍，为时间分辨FRET分析和酶反应监测提供了高灵敏度的开启发光探针。稀土对蛋白质的关键：稳定的手性稀土(III)标记与半胱氨酸残基自发和定量地反应，生成稳定的硫醚键。稀土结合标签提供了一种使用多种光谱技术探测蛋白质的工具，包括核磁共振、电子顺磁共振和时间分辨FRET实验。
A lanthanide‐binding tag site‐specifically attached to a protein presents a tool to probe the protein by multiple spectroscopic techniques, including nuclear magnetic resonance, electron paramagnetic resonance and time‐resolved luminescence spectroscopy. Here a new stable chiral LnIII tag, referred to as C12, is presented for spontaneous and quantitative reaction with a cysteine residue to generate a stable thioether bond. The synthetic protocol of the tag is relatively straightforward, and the tag is stable for storage and shipping. It displays greatly enhanced reactivity towards selenocysteine, opening a route towards selective tagging of selenocysteine in proteins containing cysteine residues. Loaded with TbIII or TmIII ions, the C12 tag readily generates pseudocontact shifts (PCS) in protein NMR spectra. It produces a relatively rigid tether between lanthanide and protein, which is beneficial for interpretation of the PCSs by single magnetic susceptibility anisotropy tensors, and it is suitable for measuring distance distributions in double electron–electron resonance experiments. Upon reaction with cysteine or other thiol compounds, the TbIII complex exhibits a 100‐fold enhancement in luminescence quantum yield, affording a highly sensitive turn‐on luminescence probe for time‐resolved FRET assays and enzyme reaction monitoring. Lanthanoid keys to proteins: A stable chiral lanthanide(III) tag reacts spontaneously and quantitatively with a cysteine residue to generate a stable thioether bond. The lanthanide‐binding tag presents a tool to probe proteins using multiple spectroscopic techniques, including NMR, electron paramagnetic resonance and time‐resolved FRET experiments.
登录
查看更多内容
使用发光灯笼型复合物在阴离子结合和传感的进步。
影响因子：
8.4
作者：
Bodman SE;Butler SJ
通讯作者：
Butler SJ
影响因子：
4.9
作者：
Gempf, Kathryn L.;Butler, Stephen J.;Parker, David
通讯作者：
Parker, David
影响因子：
4
作者：
Butler, Stephen J.;Delbianco, Martina;Zwier, Jurriaan M.
通讯作者：
Zwier, Jurriaan M.
影响因子：
15
作者：
de la Cruz, Laura;Thi Hoang Duong Nguyen;Otting, Gottfried
通讯作者：
Otting, Gottfried
影响因子：
15
作者：
Abdelkader, Elwy H.;Qianzhu, Haocheng;Otting, Gottfried
通讯作者：
Otting, Gottfried