Ligand-Induced Conformational Change of Plasmodium falciparum AMA1 Detected Using 19F NMR
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- 作者：Xiaopeng Ge ; Christopher A. MacRaild ; Shane M. Devine ; Cael O. Debono ; Geqing Wang ; Peter J. Scammells ; Martin J. Scanlon ; Robin F. Anders ; Michael Foley ; Raymond S. Norton
- 刊名：Journal of Medicinal Chemistry
- 出版年：2014
- 出版时间：August 14, 2014
- 年：2014
- 卷：57
- 期：15
- 页码：6419-6427
- 全文大小：447K
- ISSN：1520-4804
文摘
We established an efficient means of probing ligand-induced conformational change in the malaria drug target AMA1 using 19F NMR. AMA1 was labeled with 5-fluorotryptophan (5F-Trp), and the resulting 5F-Trp resonances were assigned by mutagenesis of the native Trp residues. By introducing additional Trp residues at strategic sites within a ligand-responsive loop, we detected distinct conformational consequences when various peptide and small-molecule ligands bound AMA1. Our results demonstrate an increase in flexibility in this loop caused by the native ligand, as inferred from, but not directly observed in, crystal structures. In addition, we found evidence for long-range allosteric changes in AMA1 that are not observed crystallographically. This method will be valuable in ongoing efforts to identify and characterize therapeutically relevant inhibitors of protein鈥損rotein interactions involving AMA1 and is generalizable to the study of ligand-induced conformational change in a wide range of other drug targets.