Becker Walter, Adams Luke A, Graham Bim, Wagner Gabriel E, Zangger Klaus, Otting Gottfried, Nitsche Christoph
Research School of Chemistry, Australian National University, Canberra, ACT, 2601, Australia.
Institute of Chemistry, University of Graz, 8010, Graz, Austria.
J Biomol NMR. 2018 Apr;70(4):211-218. doi: 10.1007/s10858-018-0173-6. Epub 2018 Mar 21.
Protein-ligand titrations can readily be monitored with a trimethylsilyl (TMS) tag. Owing to the intensity, narrow line shape and unique chemical shift of a TMS group, dissociation constants can be determined from straightforward 1D H-NMR spectra not only in the fast but also in the slow exchange limit. The tag is easily attached to cysteine residues and a sensitive reporter of ligand binding also at sites where it does not interfere with ligand binding or catalytic efficiency of the target protein. Its utility is demonstrated for the Zika virus NS2B-NS3 protease and the human prolyl isomerase FK506 binding protein.
蛋白质-配体滴定可以很容易地用三甲基硅烷基（TMS）标签进行监测。由于TMS基团的强度、窄线宽形状和独特的化学位移，解离常数不仅可以在快速交换极限下，而且可以在慢速交换极限下从直接的一维氢核磁共振谱中确定。该标签很容易连接到半胱氨酸残基上，并且在不干扰配体结合或靶蛋白催化效率的位点也是配体结合的灵敏报告基团。其效用已在寨卡病毒NS2B-NS3蛋白酶和人脯氨酰异构酶FK506结合蛋白中得到证明。
