DOTA-M8: An Extremely Rigid, High-Affinity Lanthanide Chelating Tag for PCS NMR Spectroscopy,Journal of the American Chemical Society

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DOTA-M8: An Extremely Rigid, High-Affinity Lanthanide Chelating Tag for PCS NMR Spectroscopy
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2009-10-21 , DOI: 10.1021/ja903233w
Daniel Häussinger ₁ , Jie-rong Huang ₁ , Stephan Grzesiek ₁

Affiliation

A new lanthanide chelating tag (M8) for paramagnetic labeling of biomolecules is presented, which is based on an eight-fold, stereoselectively methyl-substituted DOTA that can be covalently linked to the host molecule by a single disulfide bond. The steric overcrowding of the DOTA scaffold leads to an extremely rigid, kinetically and chemically inert lanthanide chelator. Its steric bulk restricts the motion of the tag relative to the host molecule. These properties result in very large pseudocontact shifts (>5 ppm) and residual dipolar couplings (>20 Hz) for Dy-M8 linked to ubiquitin, which are unprecedented for a small, single-point-attachment tag. Such large pseudocontact shifts should be well detectable even for larger proteins and distances beyond approximately 50 A. Due to its exceptionally high stability and lanthanide affinity M8 can be used under extreme chemical or physical conditions, such as those applied for protein denaturation, or when it is undesirable that buffer or protein react with excess lanthanide ions.

中文翻译：

DOTA-M8：用于 PCS NMR 光谱的极其刚性、高亲和力的镧系元素螯合标签

提出了一种用于生物分子顺磁性标记的新镧系元素螯合标签 (M8)，它基于八重立体选择性甲基取代的 DOTA，可通过单个二硫键与宿主分子共价连接。DOTA 支架的空间拥挤导致极其刚性、动力学和化学惰性的镧系元素螯合剂。它的空间体积限制了标签相对于宿主分子的运动。这些特性导致与泛素相关的 Dy-M8 产生非常大的伪接触位移 (>5 ppm) 和残余偶极耦合 (>20 Hz)，这对于小型单点连接标签而言是前所未有的。即使对于较大的蛋白质和超过大约 50 A 的距离，这种大的假接触位移也应该可以很好地检测到。

更新日期：2009-10-21

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