Molecular basis for benzodiazepine agonist action at the type 1 cholecystokinin receptor

Kaleeckal G Harikumar, Erin E Cawston, Polo C H Lam, Achyut Patil, Andrew Orry, B Henke, Ruben Abagyan, Arthur Christopoulos, Patrick Sexton, Laurence J Miller

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Understanding the molecular basis of drug action can facilitate development of more potent and selective drugs. Here, we explore the molecular basis for action of a unique small molecule ligand that is a type 1 cholecystokinin (CCK) receptor agonist and type 2 CCK receptor antagonist, GI181771X. We characterize its binding utilizing structurally-related radioiodinated ligands selective for CCK receptor subtypes that utilize the same allosteric ligand-binding pocket, using wild type receptors and chimeric constructs exchanging the distinct residues lining this pocket. Intracellular calcium assays were performed to determine biological activity. Molecular models for docking small molecule agonists to the type 1 CCK receptor were developed using a ligand-guided refinement approach. The optimal model was distinct from the previous antagonist model for the same receptor and was mechanistically consistent with the current mutagenesis data. This study revealed a key role for Leu7.39 that was predicted to interact with the isopropyl group in the N1 position of the benzodiazepine that acts as a trigger for biological activity. The molecular model was predictive of binding of other small molecule agonists, effectively distinguishing these from 1065 approved drug decoys with an AUC value of 99 . The model also selectively enriched for agonist compounds, with 130 agonists identified by ROC analysis when seeded in 2175 non-agonist ligands of the type 1 CCK receptor (AUC 78 ). Benzodiazepine agonists in this series docked in consistent pose within this pocket, with a key role played by Leu7.39, while the role of this residue was less clear for chemically-distinct agonists.
Original languageEnglish
Pages (from-to)21082 - 21095
Number of pages14
JournalJournal of Biological Chemistry
Volume288
Issue number29
DOIs
Publication statusPublished - 2013

Cite this

Harikumar, K. G., Cawston, E. E., Lam, P. C. H., Patil, A., Orry, A., Henke, B., ... Miller, L. J. (2013). Molecular basis for benzodiazepine agonist action at the type 1 cholecystokinin receptor. Journal of Biological Chemistry, 288(29), 21082 - 21095. https://doi.org/10.1074/jbc.M113.480715
Harikumar, Kaleeckal G ; Cawston, Erin E ; Lam, Polo C H ; Patil, Achyut ; Orry, Andrew ; Henke, B ; Abagyan, Ruben ; Christopoulos, Arthur ; Sexton, Patrick ; Miller, Laurence J. / Molecular basis for benzodiazepine agonist action at the type 1 cholecystokinin receptor. In: Journal of Biological Chemistry. 2013 ; Vol. 288, No. 29. pp. 21082 - 21095.
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abstract = "Understanding the molecular basis of drug action can facilitate development of more potent and selective drugs. Here, we explore the molecular basis for action of a unique small molecule ligand that is a type 1 cholecystokinin (CCK) receptor agonist and type 2 CCK receptor antagonist, GI181771X. We characterize its binding utilizing structurally-related radioiodinated ligands selective for CCK receptor subtypes that utilize the same allosteric ligand-binding pocket, using wild type receptors and chimeric constructs exchanging the distinct residues lining this pocket. Intracellular calcium assays were performed to determine biological activity. Molecular models for docking small molecule agonists to the type 1 CCK receptor were developed using a ligand-guided refinement approach. The optimal model was distinct from the previous antagonist model for the same receptor and was mechanistically consistent with the current mutagenesis data. This study revealed a key role for Leu7.39 that was predicted to interact with the isopropyl group in the N1 position of the benzodiazepine that acts as a trigger for biological activity. The molecular model was predictive of binding of other small molecule agonists, effectively distinguishing these from 1065 approved drug decoys with an AUC value of 99 . The model also selectively enriched for agonist compounds, with 130 agonists identified by ROC analysis when seeded in 2175 non-agonist ligands of the type 1 CCK receptor (AUC 78 ). Benzodiazepine agonists in this series docked in consistent pose within this pocket, with a key role played by Leu7.39, while the role of this residue was less clear for chemically-distinct agonists.",
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Harikumar, KG, Cawston, EE, Lam, PCH, Patil, A, Orry, A, Henke, B, Abagyan, R, Christopoulos, A, Sexton, P & Miller, LJ 2013, 'Molecular basis for benzodiazepine agonist action at the type 1 cholecystokinin receptor' Journal of Biological Chemistry, vol. 288, no. 29, pp. 21082 - 21095. https://doi.org/10.1074/jbc.M113.480715

Molecular basis for benzodiazepine agonist action at the type 1 cholecystokinin receptor. / Harikumar, Kaleeckal G; Cawston, Erin E; Lam, Polo C H; Patil, Achyut; Orry, Andrew; Henke, B; Abagyan, Ruben; Christopoulos, Arthur; Sexton, Patrick; Miller, Laurence J.

In: Journal of Biological Chemistry, Vol. 288, No. 29, 2013, p. 21082 - 21095.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Molecular basis for benzodiazepine agonist action at the type 1 cholecystokinin receptor

AU - Harikumar, Kaleeckal G

AU - Cawston, Erin E

AU - Lam, Polo C H

AU - Patil, Achyut

AU - Orry, Andrew

AU - Henke, B

AU - Abagyan, Ruben

AU - Christopoulos, Arthur

AU - Sexton, Patrick

AU - Miller, Laurence J

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N2 - Understanding the molecular basis of drug action can facilitate development of more potent and selective drugs. Here, we explore the molecular basis for action of a unique small molecule ligand that is a type 1 cholecystokinin (CCK) receptor agonist and type 2 CCK receptor antagonist, GI181771X. We characterize its binding utilizing structurally-related radioiodinated ligands selective for CCK receptor subtypes that utilize the same allosteric ligand-binding pocket, using wild type receptors and chimeric constructs exchanging the distinct residues lining this pocket. Intracellular calcium assays were performed to determine biological activity. Molecular models for docking small molecule agonists to the type 1 CCK receptor were developed using a ligand-guided refinement approach. The optimal model was distinct from the previous antagonist model for the same receptor and was mechanistically consistent with the current mutagenesis data. This study revealed a key role for Leu7.39 that was predicted to interact with the isopropyl group in the N1 position of the benzodiazepine that acts as a trigger for biological activity. The molecular model was predictive of binding of other small molecule agonists, effectively distinguishing these from 1065 approved drug decoys with an AUC value of 99 . The model also selectively enriched for agonist compounds, with 130 agonists identified by ROC analysis when seeded in 2175 non-agonist ligands of the type 1 CCK receptor (AUC 78 ). Benzodiazepine agonists in this series docked in consistent pose within this pocket, with a key role played by Leu7.39, while the role of this residue was less clear for chemically-distinct agonists.

AB - Understanding the molecular basis of drug action can facilitate development of more potent and selective drugs. Here, we explore the molecular basis for action of a unique small molecule ligand that is a type 1 cholecystokinin (CCK) receptor agonist and type 2 CCK receptor antagonist, GI181771X. We characterize its binding utilizing structurally-related radioiodinated ligands selective for CCK receptor subtypes that utilize the same allosteric ligand-binding pocket, using wild type receptors and chimeric constructs exchanging the distinct residues lining this pocket. Intracellular calcium assays were performed to determine biological activity. Molecular models for docking small molecule agonists to the type 1 CCK receptor were developed using a ligand-guided refinement approach. The optimal model was distinct from the previous antagonist model for the same receptor and was mechanistically consistent with the current mutagenesis data. This study revealed a key role for Leu7.39 that was predicted to interact with the isopropyl group in the N1 position of the benzodiazepine that acts as a trigger for biological activity. The molecular model was predictive of binding of other small molecule agonists, effectively distinguishing these from 1065 approved drug decoys with an AUC value of 99 . The model also selectively enriched for agonist compounds, with 130 agonists identified by ROC analysis when seeded in 2175 non-agonist ligands of the type 1 CCK receptor (AUC 78 ). Benzodiazepine agonists in this series docked in consistent pose within this pocket, with a key role played by Leu7.39, while the role of this residue was less clear for chemically-distinct agonists.

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DO - 10.1074/jbc.M113.480715

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JF - Journal of Biological Chemistry

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