Solution structure ensemble of human obesity-associated protein FTO reveals druggable surface pockets at the interface between the N- and C-terminal domain

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Khatiwada, Balabhadra
Nguyen, Trang T.
Purslow, Jeffrey A.
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Elsevier Inc.
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ChemistryBiochemistry, Biophysics and Molecular Biology
The fat mass and obesity-associated FTO protein catalyzes demethylation of the N6-methyladenosine, an epigenetic mark that controls several metabolic pathways by modulating the transcription, translation, and cellular localization of RNA molecules. Since the discovery that its overexpression links to the development of obesity and cancer, FTO was the target of screening campaigns and structure-based drug design efforts. Although several FTO inhibitors were generated, these often lack potency or selectivity. Herein, we investigate the structure and dynamics of human FTO in solution. We show that the structure of the catalytic N-terminal domain is unstable in the absence of the C-terminal domain, which explains why the isolated N-terminal domain is incompetent for catalysis and suggests that the domain interaction represents a target for the development of specific inhibitors. Then, by using NMR relaxation measurements, we show that the interface between the FTO structural domains, the active site, and several peripheral loops undergo conformational dynamics on both the picosecond–nanosecond and microsecond–millisecond timescales. Consistent with this, we found that the backbone amide residual dipolar couplings measured for FTO in phage pf1 are inconsistent with the static crystal structure of the enzyme. Finally, we generated a conformational ensemble for apo FTO that satisfies the solution NMR data by combining the experimental residual dipolar couplings with accelerated molecular dynamics simulations. Altogether, the structural ensemble reported in this work provides an atomic-resolution model of apo FTO and reveals transient surface pockets at the domain interface that represent potential targets for the design of allosteric inhibitors.
This article is published as Khatiwada, Balabhadra, Trang T. Nguyen, Jeffrey A. Purslow, and Vincenzo Venditti. "Solution structure ensemble of human obesity-associated protein FTO reveals druggable surface pockets at the interface between the N-and C-terminal domain." Journal of Biological Chemistry 298, no. 5 (2022): 101907. DOI: 10.1016/j.jbc.2022.101907. Copyright 2022 The Authors. Attribution 4.0 International (CC BY 4.0). Posted with permission.