한국자기공명학회논문지 (Journal of the Korean Magnetic Resonance Society)

한국자기공명학회 (Korean Magnetic Resonance Society)
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NMR-based Analysis of the Interaction between Pomalidomide and the C-terminal Domain of Cereblon

  • Bhattarai Shishir (College of Pharmacy and Gachon Institute of Pharmaceutical Sciences, Gachon University) ;
  • Sung Jean Park (College of Pharmacy and Gachon Institute of Pharmaceutical Sciences, Gachon University)
  • 투고 : 2025.06.26
  • 심사 : 2025.08.05
  • 발행 : 2025.09.20
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초록

Cereblon (CRBN) is a substrate receptor of the CRL4 E3 ubiquitin ligase complex and a key component in the mechanism of action of immunomodulatory drugs (IMiDs) such as pomalidomide. In this study, we investigated the molecular interaction between pomalidomide and the C-terminal thalidomide-binding domain of CRBN (CdCRBN) using solution NMR spectroscopy, isothermal titration calorimetry (ITC), and fluorescence-based competition assays. Two glycine residues, G354 and G383, located near the ligand-binding site, were employed as markers to monitor binding-induced chemical shift perturbations. The 1H, 15N-HSQC titration revealed concentration-dependent chemical shift changes, indicating fast exchange kinetics. The dissociation constant (Kd) calculated by NMR was 55 ± 1.8 µM, while ITC analysis showed a stronger binding affinity (Kd = 12.5 µM), suggesting an enthalpy-driven interaction. A fluorescence resonance energy transfer (FRET)-based competition assay using MANT-uracil further supported the interaction, yielding a Ki of 2.1 µM. These results confirm that pomalidomide binds to the canonical thalidomide-binding pocket of CdCRBN, providing a structural and thermodynamic basis for its function as a molecular glue.

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과제정보

This work was supported by the Gachon University research fund of 2023 (GCU-202300420001).This research was also supported by the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology (2020R1A6A1A0304370812).

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