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Molecular basis for cross-activation of NPFF2R by a short PrRP-related peptide

Acta Pharmacologica Sinica (2026)Cite this article

Abstract

Prolactin-releasing peptide (PrRP) is an endogenous ligand for the PrRPR, whose activation has been linked to anti-obesity effects. However, PrRP and its analogs also activate the neuropeptide FF receptor 2 (NPFF2R), which is associated with adverse cardiovascular effects. Understanding how PrRP-related peptides differentially engage these two distinct receptors is critical for developing safer, more selective therapeutics. In this study, we present cryo-EM structures of the PrRP analog GUB08248 bound to PrRPR-Gαq and NPFF2R-Gαi at resolutions of 2.45 Å and 2.85 Å, respectively. These structures reveal a conserved ligand recognition mode across both receptors, while highlighting distinct receptor-specific interactions. The NPFF2R-Gαi complex further uncovers key features of receptor activation and G protein coupling. Together, our results offer structural insights that could guide structure-based drug design strategies favoring PrRPR selectivity, thereby advancing the therapeutic potential of the PrRP-PrRPR axis for obesity treatment.

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Fig. 1: Cryo-EM structures of Gαq-coupled PrRPR and Gαi-coupled NPFF2R in complex with GUB08248.
Fig. 2: Molecular basis of GUB08248 recognition by PrRPR.
Fig. 3: Molecular basis of GUB08248 recognition by NPFF2R.
Fig. 4: Comparison of the GUB08248 binding mode between PrRPR and NPFF2R.
Fig. 5: Conformational changes in NPFF2R upon activation by GUB08248.
Fig. 6: G protein coupling of NPFF2R.

Data availability

The atomic coordinates and the electron microscopy maps have been deposited in the Protein Data Bank (PDB) and the Electron Microscopy Data Bank (EMDB) under accession codes 9V0X and EMD-64674 (composite map), EMD-64675 (receptor-locally refined map), and EMD-64678 (consensus map) for the GUB08248-PrRPR-Gαq complex, and 9V1H and EMD-64686 (consensus map), EMD-64690 (receptor-locally refined map), and EMD-64693 (composite map) for the GUB08248-NPFF2R-Gαi complex.

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Acknowledgements

The cryo-EM data were collected at the Advanced Center for Electron Microscopy at Shanghai Institute of Materia Medica, Chinese Academy of Sciences. This work was supported by National Natural Science Foundation of China (32371255 and 32071203 to LHZ, 82495184, 32130022 and 82121005 to HEX, 82404881 to QNY); Natural Science Foundation of Shanghai (23ZR1475200 to LHZ); the National Key R&D Program of China (2022YFC2703105 to HEX); CAS Strategic Priority Research Program (XDB37030103 to HEX); Shanghai Municipal Science and Technology Major Project (2019SHZDZX02 to HEX); the Young Innovator Association of CAS (Y2022078 to LHZ); the Lingang Laboratory (LG-GG-202204-01 to HEX); State Key Laboratory of Drug Research (SKLDR-2023-TT-04 to HEX).

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  1. These authors contributed equally: Xin Li, Shuai Li.

Authors and Affiliations

  1. State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China

    Xin Li, Shuai Li, Hong Shan, Qian He, Min Zhang, Yang Li, Wen Hu, Kai Wu & H. Eric Xu

  2. University of Chinese Academy of Sciences, Beijing, 100049, China

    Xin Li, Qian He, Min Zhang & H. Eric Xu

  3. State Key Laboratory of Medical Genomics, Research Center for Medicinal Structural Biology, National Research Center for Translational Medicine at Shanghai, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China

    Qing-ning Yuan, H. Eric Xu & Li-hua Zhao

  4. Lingang Laboratory, Shanghai, 201100, China

    Xin-heng He

  5. School of Pharmacy, Fudan University, Shanghai, 201203, China

    Yang Li

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Contributions

XL screened the expression constructs, purified proteins, and prepared samples for structural determination, conducted structural analysis, carried out functional assays, generated figures, and drafted the initial manuscript; SL and QNY performed cryo-EM data processing, model building, and structure refinement; QH and MZ prepared cryo-EM grids; HS and XHH conducted the computational studies; YL contributed to parts of surface expression analysis; WH and KW assisted with data collection; LHZ and HEX conceived and supervised the project and revised the manuscript.

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Correspondence to H. Eric Xu or Li-hua Zhao.

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Prof H. Eric Xu is one of the Associate Editors of the journal and was not involved in the peer review or the decision making of the article. The authors declare no other competing interests.

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Li, X., Li, S., Shan, H. et al. Molecular basis for cross-activation of NPFF2R by a short PrRP-related peptide. Acta Pharmacol Sin (2026). https://doi.org/10.1038/s41401-025-01741-1

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