9JG0

Cryo-EM structure of neuropeptide FF receptor 2 in the ligand-free state with BRIL fusion, anti-BRIL Fab, and nanobody

Summary for 9JG0

• Entry DOI: 10.2210/pdb9jg0/pdb
• EMDB information: 61446
• Descriptor: Isoform 2 of Neuropeptide FF receptor 2,Soluble cytochrome b562, Anti-BRIL fab heavy chain, Anti-fab nanobody, ... (4 entities in total)
• Functional Keywords: gpcr, membrane protein
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 4
• Total formula weight: 122737.14

Authors

Kim, J.,Choi, H.-J. (deposition date: 2024-09-05, release date: 2025-04-09)

Primary citation

Kim, J.,Hong, S.,Lee, H.,Lee, H.S.,Park, C.,Kim, J.,Im, W.,Choi, H.J.
Structural insights into the selective recognition of RF-amide peptides by neuropeptide FF receptor 2.
Embo Rep., 2025

PubMed Abstract: Neuropeptide FF Receptor 2 (NPFFR2), a G-protein-coupled receptor, plays a role in pain modulation and diet-induced thermogenesis. While NPFFR2 is strongly activated by neuropeptides FF (NPFFs), it shows low activity in response to RF-amide-related peptides (RFRPs), despite the peptides belonging to a shared family. In contrast, NPFFR1, which shares high sequence similarity with NPFFR2, is activated by RFRPs and regulates reproductive hormone balance. The molecular basis for these receptor-specific interactions with their RF-amide peptides remains unclear. Here, we present cryo-electron microscopy structures of NPFFR2 in its active state bound to the agonist RF-amide peptide hNPSF, and in its ligand-free state. Structural analysis reveals that the C-terminal RF-amide moiety engages conserved residues in the transmembrane domain, while the N-terminal segment interacts in a receptor subtype-specific manner. Key selectivity-determining residues in NPFFR2 are also identified. A homology model of NPFFR1 bound to RFRP, supported by mutagenesis studies, further validates this selectivity mechanism. Additionally, structural comparison between the inactive and active states of NPFFR2 suggests a TM3-mediated activation mechanism. These findings provide insights into RF-amide peptide recognition by NPFF receptors.

PubMed: 40128413
DOI: 10.1038/s44319-025-00428-2

Experimental method

ELECTRON MICROSCOPY (2.91 Å)