Abstract
FROUNT is a cytoplasmic protein that interacts with the membrane-proximal C-terminal regions (Pro-Cs) of the CCR2 and CCR5 chemokine receptors. The interactions between FROUNT and the chemokine receptors play an important role in the migration of inflammatory immune cells. Therefore, FROUNT is a potential drug target for inflammatory diseases. However, the structural basis of the interactions between FROUNT and the chemokine receptors remains to be elucidated. We previously identified the C-terminal region (residues 532–656) of FROUNT as the structural domain responsible for the Pro-C binding, referred to as the chemokine receptor-binding domain (CRBD), and then constructed its mutant, bearing L538E/P612S mutations, with improved NMR spectral quality, referred to as CRBD_LEPS. We now report the main-chain and side-chain 1H, 13C, and 15N resonance assignments of CRBD_LEPS. The NMR signals of CRBD_LEPS were well dispersed and their intensities were uniform on the 1H–15N HSQC spectrum, and thus almost all of the main-chain and side-chain resonances were assigned. This assignment information provides the foundation for NMR studies of the three-dimensional structure of CRBD_LEPS in solution and its interactions with chemokine receptors.
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Acknowledgements
We gratefully acknowledge M. Yokochi and F. Inagaki for NMR analyses. This work was supported in part by the Targeted Proteins Research Program (TPRP) from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT) (to S.Y., E.T., Y.T., K.M. and H.T.), by Practical Research for Innovative Cancer Control from the Japan Agency for Medical Research and Development (AMED) (to S.Y., M.T., E.T., Y.T., K.M. and H.T.), by Project for Development of Innovative Research on Cancer Therapeutics (P-DIRECT) from AMED (to K.M.), by Project for Cancer Research and Therapeutic Evolution (P-CREATE) from AMED (to S.Y., M.T., Y.T., K.M. and H.T.), by a Grant-in-Aid for Young Scientists (B) (JP19790064) from MEXT (to S.Y.), by the KUMAYAKU Alumni Research Fund (to S.Y.), and by the Adaptable and Seamless Technology Transfer Program through target-driven R&D (A-STEP), Japan Science and Technology Agency (JST) (to H.T.).
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Yoshinaga, S., Ishida, N., Tsuji, T. et al. 1H, 13C and 15N resonance assignments for a chemokine receptor-binding domain of FROUNT, a cytoplasmic regulator of chemotaxis. Biomol NMR Assign 12, 259–262 (2018). https://doi.org/10.1007/s12104-018-9819-2
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DOI: https://doi.org/10.1007/s12104-018-9819-2