19F-NMR Unveils the Ligand-Induced Conformation of a Catalytically Inactive Twisted Homodimer of tRNA–Guanine Transglycosylase

Nguyen, Andreas; Gemmecker, Gerd; Softley, Charlotte A.; Movsisyan, Levon D.; Pfaffeneder, Toni; Heine, Andreas; Reuter, Klaus; Diederich, François; Sattler, Michael; Klebe, Gerhard

Abstract

Understanding the structural arrangements of protein oligomers can support the design of ligands that interfere with their function in order to develop new therapeutic concepts for disease treatment. Recent crystallographic studies have elucidated a novel twisted and functionally inactive form of the homodimeric enzyme tRNA–guanine transglycosylase (TGT), a putative target in the fight against shigellosis. Active-site ligands have been identified that stimulate the rearrangement of one monomeric subunit by 130° against the other one to form an inactive twisted homodimer state. To assess whether the crystallographic observations also reflect the conformation in solution and rule out effects from crystal packing, we performed 19F-NMR spectroscopy with the introduction of 5-fluorotryptophans at four sites in TGT. The inhibitor-induced conformation of TGT in solution was assessed based on 19F-NMR chemical shift perturbations. We investigated the effect of C(4) substituted lin-benzoguanine ligands and identified a correlation between dynamic protein rearrangements and ligand-binding features in the corresponding crystal structures. These involve the destabilization of a helix next to the active site and the integrity of a flexible loop–helix motif. Ligands that either completely lack an attached C(4) substituent or use it to stabilize the geometry of the functionally competent dimer state do not indicate the presence of the twisted dimer form in the NMR spectra. The perturbation of crucial structural motifs in the inhibitors correlates with an increasing formation of the inactive twisted dimer state, suggesting these ligands are able to shift a conformational equilibrium from active C2-symmetric to inactive twisted dimer conformations. These findings suggest a novel concept for the design of drug candidates for further development.

Citation

Nguyen, A., Gemmecker, G., Softley, C. A., Movsisyan, L. D., Pfaffeneder, T., Heine, A., …Klebe, G. (2022). 19F-NMR Unveils the Ligand-Induced Conformation of a Catalytically Inactive Twisted Homodimer of tRNA–Guanine Transglycosylase. ACS chemical biology, 17(7), 1745-1755. https://doi.org/10.1021/acschembio.2c00080

• Journal Article Type: Article
• Acceptance Date: Jun 9, 2022
• Online Publication Date: Jun 28, 2022
• Publication Date: Jul 15, 2022
• Deposit Date: Feb 24, 2025
• Journal: ACS Chemical Biology
• Print ISSN: 1554-8929
• Electronic ISSN: 1554-8937
• Publisher: American Chemical Society
• Peer Reviewed: Peer Reviewed
• Volume: 17
• Issue: 7
• Pages: 1745-1755
• DOI: https://doi.org/10.1021/acschembio.2c00080
• Public URL: https://keele-repository.worktribe.com/output/1078786
• Publisher URL: https://pubs.acs.org/doi/10.1021/acschembio.2c00080