Ligand-induced conformational change of Plasmodium falciparum AMA1 detected using 19F NMR

Xiaopeng Ge, Christopher Andrew MacRaild, Shane Devine, Cael Debono, Geqing Wang, Peter John Scammells, Martin Scanlon, Robin F Anders, Michael Foley, Raymond Stanley Norton

Research output: Contribution to journalArticleResearchpeer-review

Abstract

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-protein interactions involving AMA1 and is generalizable to the study of ligand-induced conformational change in a wide range of other drug targets.
Original languageEnglish
Pages (from-to)6419 - 6427
Number of pages9
JournalJournal of Medicinal Chemistry
Volume57
Issue number15
DOIs
Publication statusPublished - 2014

Cite this

@article{81eed6e4e7df49c5955c265add778f5d,
title = "Ligand-induced conformational change of Plasmodium falciparum AMA1 detected using 19F NMR",
abstract = "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-protein interactions involving AMA1 and is generalizable to the study of ligand-induced conformational change in a wide range of other drug targets.",
author = "Xiaopeng Ge and MacRaild, {Christopher Andrew} and Shane Devine and Cael Debono and Geqing Wang and Scammells, {Peter John} and Martin Scanlon and Anders, {Robin F} and Michael Foley and Norton, {Raymond Stanley}",
year = "2014",
doi = "10.1021/jm500390g",
language = "English",
volume = "57",
pages = "6419 -- 6427",
journal = "Journal of Medicinal Chemistry",
issn = "0022-2623",
publisher = "AMER CHEMICAL SOC",
number = "15",

}

Ligand-induced conformational change of Plasmodium falciparum AMA1 detected using 19F NMR. / Ge, Xiaopeng; MacRaild, Christopher Andrew; Devine, Shane; Debono, Cael; Wang, Geqing; Scammells, Peter John; Scanlon, Martin; Anders, Robin F; Foley, Michael; Norton, Raymond Stanley.

In: Journal of Medicinal Chemistry, Vol. 57, No. 15, 2014, p. 6419 - 6427.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Ligand-induced conformational change of Plasmodium falciparum AMA1 detected using 19F NMR

AU - Ge, Xiaopeng

AU - MacRaild, Christopher Andrew

AU - Devine, Shane

AU - Debono, Cael

AU - Wang, Geqing

AU - Scammells, Peter John

AU - Scanlon, Martin

AU - Anders, Robin F

AU - Foley, Michael

AU - Norton, Raymond Stanley

PY - 2014

Y1 - 2014

N2 - 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-protein interactions involving AMA1 and is generalizable to the study of ligand-induced conformational change in a wide range of other drug targets.

AB - 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-protein interactions involving AMA1 and is generalizable to the study of ligand-induced conformational change in a wide range of other drug targets.

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