Abstract
Apical membrane antigen 1 (AMA1) is essential for the invasion of host cells by malaria parasites. Several small-molecule ligands have been shown to bind to a conserved hydrophobic cleft in Plasmodium falciparum AMA1. However, a lack of detailed structural information on the binding pose of these molecules has hindered their further optimisation as inhibitors. We have developed a spin-labelled peptide based on RON2, the native binding partner of AMA1, to probe the binding sites of compounds on PfAMA1. The crystal structure of this peptide bound to PfAMA1 shows that it binds at one end of the hydrophobic groove, leaving much of the binding site unoccupied and allowing fragment hits to bind without interference. In paramagnetic relaxation enhancement (PRE)-based NMR screening, the 1 H relaxation rates of compounds binding close to the probe were enhanced. Compounds experienced different degrees of PRE as a result of their different orientations relative to the spin label while bound to AMA1. Thus, PRE-derived distance constraints can be used to identify binding sites and guide further hit optimisation.
| Original language | English |
|---|---|
| Pages (from-to) | 603-612 |
| Number of pages | 10 |
| Journal | ChemMedChem |
| Volume | 14 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 5 Mar 2019 |
Keywords
- AMA1
- drug discovery
- malaria
- NMR spectroscopy
- spin label
Cite this
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Identification of the Binding Site of Apical Membrane Antigen 1 (AMA1) Inhibitors Using a Paramagnetic Probe. / Akter, Mansura; Drinkwater, Nyssa; Devine, Shane M.; Drew, Simon C.; Krishnarjuna, Bankala; Debono, Cael O.; Wang, Geqing; Scanlon, Martin J.; Scammells, Peter J.; McGowan, Sheena; MacRaild, Christopher A.; Norton, Raymond S.
In: ChemMedChem, Vol. 14, No. 5, 05.03.2019, p. 603-612.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Identification of the Binding Site of Apical Membrane Antigen 1 (AMA1) Inhibitors Using a Paramagnetic Probe
AU - Akter, Mansura
AU - Drinkwater, Nyssa
AU - Devine, Shane M.
AU - Drew, Simon C.
AU - Krishnarjuna, Bankala
AU - Debono, Cael O.
AU - Wang, Geqing
AU - Scanlon, Martin J.
AU - Scammells, Peter J.
AU - McGowan, Sheena
AU - MacRaild, Christopher A.
AU - Norton, Raymond S.
PY - 2019/3/5
Y1 - 2019/3/5
N2 - Apical membrane antigen 1 (AMA1) is essential for the invasion of host cells by malaria parasites. Several small-molecule ligands have been shown to bind to a conserved hydrophobic cleft in Plasmodium falciparum AMA1. However, a lack of detailed structural information on the binding pose of these molecules has hindered their further optimisation as inhibitors. We have developed a spin-labelled peptide based on RON2, the native binding partner of AMA1, to probe the binding sites of compounds on PfAMA1. The crystal structure of this peptide bound to PfAMA1 shows that it binds at one end of the hydrophobic groove, leaving much of the binding site unoccupied and allowing fragment hits to bind without interference. In paramagnetic relaxation enhancement (PRE)-based NMR screening, the 1 H relaxation rates of compounds binding close to the probe were enhanced. Compounds experienced different degrees of PRE as a result of their different orientations relative to the spin label while bound to AMA1. Thus, PRE-derived distance constraints can be used to identify binding sites and guide further hit optimisation.
AB - Apical membrane antigen 1 (AMA1) is essential for the invasion of host cells by malaria parasites. Several small-molecule ligands have been shown to bind to a conserved hydrophobic cleft in Plasmodium falciparum AMA1. However, a lack of detailed structural information on the binding pose of these molecules has hindered their further optimisation as inhibitors. We have developed a spin-labelled peptide based on RON2, the native binding partner of AMA1, to probe the binding sites of compounds on PfAMA1. The crystal structure of this peptide bound to PfAMA1 shows that it binds at one end of the hydrophobic groove, leaving much of the binding site unoccupied and allowing fragment hits to bind without interference. In paramagnetic relaxation enhancement (PRE)-based NMR screening, the 1 H relaxation rates of compounds binding close to the probe were enhanced. Compounds experienced different degrees of PRE as a result of their different orientations relative to the spin label while bound to AMA1. Thus, PRE-derived distance constraints can be used to identify binding sites and guide further hit optimisation.
KW - AMA1
KW - drug discovery
KW - malaria
KW - NMR spectroscopy
KW - spin label
UR - http://www.scopus.com/inward/record.url?scp=85061638809&partnerID=8YFLogxK
U2 - 10.1002/cmdc.201800802
DO - 10.1002/cmdc.201800802
M3 - Article
VL - 14
SP - 603
EP - 612
JO - ChemMedChem
JF - ChemMedChem
SN - 1860-7179
IS - 5
ER -