Journal article
Developing inhibitors of the guanosine triphosphate hydrolysis accelerating activity of Regulator of G protein Signaling-14
The Journal of biological chemistry, Vol.301(10), 110611
10/2025
DOI: 10.1016/j.jbc.2025.110611
PMCID: PMC12495443
PMID: 40848973
Abstract
Regulator of G protein Signaling-14 (RGS14), an intracellular inactivator of G protein-coupled receptor (GPCR) signaling, is considered an undruggable protein given its shallow and relatively featureless protein-protein interaction interface combined with a distal allosteric site prone to nonspecific inhibition by thiol-reactive compounds. Here, we identify and validate a tractable chemotype that selectively and non-covalently inhibits RGS14 GTPase-accelerating protein (GAP) activity. Combining structure-guided virtual screening, ligand docking across multiple receptor conformers, and enrichment validation, we progressed from a first-generation active, Z90276197, to over 40 second-generation analogs with improved potency. These inhibitors are predicted to engage the solvent-exposed “canyon” in the RGS14 RGS-box that interacts with the Gα switch I region. Binding pose predictions underscored the importance of non-polar interactions and shape complementarity over polar interactions in engaging this Gα-binding canyon and revealed an “ambidextrous” pattern of R1- and R2-group orientations. GAP inhibition was confirmed in fluorescence-based and gold-standard radioactive GTP hydrolysis assays. Two second-generation analogs, Z55660043 and Z55627844, inhibited RGS14 GAP activity in both assays and without measurable cytotoxicity. Deep learning-based scoring of predicted docking poses further supported observed affinity gains from R3-group additions. One analog demonstrated favorable in vivo pharmacokinetics and CNS penetration. Collectively, our findings establish tractable, non-covalent, small molecule inhibition of a G protein regulatory interface and illustrate how machine learning-enhanced docking can guide ligand optimization for shallow protein surfaces. This work opens the door to future development of RGS14 inhibitors as potential therapeutics for central nervous system and metabolic disorders.
Details
- Title: Subtitle
- Developing inhibitors of the guanosine triphosphate hydrolysis accelerating activity of Regulator of G protein Signaling-14
- Creators
- Percy S. Agogo-Mawuli - University of North Texas Health Science CenterIsra Sadiya - University of HaifaTigran M. Abramyan - Atomwise Inc., San Francisco, CADustin E. Bosch - University of IowaKyle A. Emmitte - University of North Texas Health Science CenterLuis M. Colón-Pérez - University of North Texas Health Science CenterMickey Kosloff - University of HaifaDavid P. Siderovski - University of North Texas Health Science Center
- Resource Type
- Journal article
- Publication Details
- The Journal of biological chemistry, Vol.301(10), 110611
- DOI
- 10.1016/j.jbc.2025.110611
- PMID
- 40848973
- PMCID
- PMC12495443
- NLM abbreviation
- J Biol Chem
- ISSN
- 0021-9258
- eISSN
- 1083-351X
- Publisher
- Elsevier Inc
- Grant note
- US National Institute on Drug Abuse: DA048153 HSC Division of Research and InnovationIsrael Science Foundation (ISF)Azrieli Foundation: 3512/19 Council for Higher Education through the Data Science Research Center at the University of HaifaHSC Presidential Endowment
Work was supported in part by the US National Institute on Drug Abuse (R01 grant DA048153, to D. P. S) , a Team Science award from the HSC Division of Research and Innovation (to K. A. E., L. C.-P., and D. P. S.) , the Israel Science Foundation (ISF) and the Azrieli Foundation (grant 3512/19 to M. K.) , a grant from the Council for Higher Education through the Data Science Research Center at the University of Haifa (to M. K.) , and an HSC Presidential Endowment to the Chair of Pharmacology and Neuroscience (to D. P. S.) .
- Language
- English
- Electronic publication date
- 08/2025
- Date published
- 10/2025
- Academic Unit
- Pathology
- Record Identifier
- 9984949225802771
Metrics
2 Record Views