Chen Wan-Na, Otting Gottfried
Research School of Chemistry, Australian National University, Canberra, ACT 2601, Australia.
College of Pharmacy, Jinan University, Guangzhou, Guangdong 510632, China.
ACS Med Chem Lett. 2018 Jan 12;9(2):109-113. doi: 10.1021/acsmedchemlett.7b00464. eCollection 2018 Feb 8.
Few methods allow determining the binding site of tightly binding ligands. We show that ligands containing a -butyl (e.g., Boc) group produce easily observable nuclear Overhauser effects (NOE) with the target protein even when the -butyl group is not highly solvent exposed. NOEs with methyl groups of the target protein are readily assigned by selectively isotope labeling, presenting a practical and quick way to pinpoint the location of the ligand without any prior specific nuclear magnetic resonance assignments of the protein. The approach works for nonexchanging ligands as well as for weakly binding ligands.
很少有方法能够确定紧密结合配体的结合位点。我们发现，即使叔丁基（如Boc基团）并未高度暴露于溶剂中，含有叔丁基的配体与目标蛋白也会产生易于观察到的核Overhauser效应（NOE）。通过选择性同位素标记，可以轻松确定目标蛋白甲基基团的NOE，从而提供了一种实用且快速的方法来确定配体的位置，而无需事先对蛋白质进行任何特定的核磁共振归属。该方法适用于非交换配体以及弱结合配体。
