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Related Experiment Video

Updated: May 7, 2026

Strategic Screening and Characterization of the Visual GPCR-mini-G Protein Signaling Complex for Successful Crystallization
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Decoding the structure of GPR151 via NELiS.

Yumeng Wang1, Luyu Fan2, Qianqian Song1

  • 1Key Laboratory of Multi-Cell Systems, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.

Proceedings of the National Academy of Sciences of the United States of America
|May 5, 2026
PubMed
Summary
This summary is machine-generated.

A new method, Nb6-Enabled Ligand-Free Stabilization Platform (NELiS), stabilizes orphan G protein-coupled receptors (GPCRs) without ligands. This breakthrough enables structural and functional studies of previously intractable receptors like GPR151.

Keywords:
GPCRGPR151orphan receptor

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Area of Science:

  • Structural Biology
  • Neuroscience
  • Pharmacology

Background:

  • Orphan G protein-coupled receptors (GPCRs) present significant challenges for structural and functional studies due to the absence of known ligands and inherent instability.
  • Conventional methods for assessing protein stability and facilitating purification are often ligand-dependent, limiting their application to orphan GPCRs.

Purpose of the Study:

  • To develop a novel ligand- and purification-independent method for identifying stabilizing mutations in orphan GPCRs.
  • To investigate the structure and function of the orphan GPR151 receptor, implicated in neuropsychiatric disorders.

Main Methods:

  • Development and application of the Nb6-Enabled Ligand-Free Stabilization Platform (NELiS) to identify stabilizing mutations.
  • Purification of stabilized GPR151 and subsequent nanobody discovery for structural determination.
  • X-ray crystallography and functional assays to elucidate receptor structure and mechanism.

Main Results:

  • NELiS successfully identified four mutations that enhanced the thermostability and expression of GPR151, enabling its purification.
  • Structural analysis revealed unique activation-resistant features and an autoinhibitory N-terminal region in GPR151.
  • Functional studies confirmed an unconventional activation mechanism and the essential role of the N terminus in GPR151 maturation and trafficking.

Conclusions:

  • The NELiS platform is a generalizable tool for overcoming stabilization challenges in orphan GPCR research.
  • The study provides critical structural and functional insights into GPR151, a potential therapeutic target for neuropsychiatric disorders.