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Extracellular nanobody screening using conformationally stable GPCR variants
Xin Zhang1,2, Kaixuan Gao1,2, Jia Nie3
1State Key Laboratory of Membrane Biology, Tsinghua-Peking Center for Life Sciences, School of Pharmaceutical Sciences, Tsinghua University, Beijing 100084, China.
Summary
Researchers developed a universal method to stabilize G protein-coupled receptors (GPCRs) in specific conformations. This technique aids in drug discovery by providing stable receptor targets for screening potential therapeutics.
Area of Science:
- Biochemistry
- Structural Biology
- Pharmacology
Background:
- G protein-coupled receptors (GPCRs) are crucial drug targets, but their dynamic nature complicates ligand screening.
- Obtaining stable, purified GPCRs in specific conformations, especially active states, is a significant challenge in drug development.
Purpose of the Study:
- To develop a universal strategy for stabilizing GPCRs in desired conformations for enhanced drug screening.
- To create tools for identifying conformation-specific ligands, improving drug discovery efficiency and specificity.
Main Methods:
- De novo design of a fusion protein to stabilize the M1 muscarinic acetylcholine receptor (M1R) in its active conformation.
- Application of the stabilization strategy to other GPCRs to demonstrate generalizability.
- Screening of a nanobody library against stabilized active and inactive M1R states using yeast display technology.
Main Results:
- Successfully stabilized M1R in its active conformation using the designed fusion protein.
- Demonstrated the generalizability of the stabilization approach across different GPCRs.
- Identified conformation-specific nanobodies that recognize distinct M1R states.
Conclusions:
- The developed universal approach effectively stabilizes GPCRs in specific conformations, overcoming a major hurdle in drug discovery.
- The stabilized GPCRs and identified nanobodies serve as valuable tools for developing more selective therapeutic agents.
- This method significantly enhances the efficiency and specificity of GPCR-targeted drug discovery.
Related Concept Videos
01:21G-protein Coupled Receptors
G-protein coupled receptors are ligand binding receptors that indirectly affect changes in the cell. The actual receptor is a single polypeptide that transverses the cell membrane seven times creating intracellular and extracellular loops. The extracellular loops create a ligand specific pocket which binds to neurotransmitters or hormones. The intracellular loops holds onto the G-protein.
01:21G-protein Coupled Receptors
G-protein coupled receptors are ligand binding receptors that indirectly affect changes in the cell. The actual receptor is a single polypeptide that transverses the cell membrane seven times creating intracellular and extracellular loops. The extracellular loops create a ligand specific pocket which binds to neurotransmitters or hormones. The intracellular loops holds onto the G-protein.
01:15G Protein-coupled Receptors
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GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...
GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...
01:12GPCR Desensitization
G protein-coupled receptor (GPCR) signaling plays a crucial role in cell functioning. GPCR desensitization is an equally essential process. It allows cells to respond to changing environments and regain sensitivity to new stimuli while preventing unnecessary stimulation when no longer needed. Prolonged exposure to stimuli leads to GPCR desensitization. It involves blocking the receptors from binding and activating additional G proteins. This inhibits activation of downstream effectors, thereby...
01:09GPCRs Regulate Adenylyl Cylase Activity
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01:30Transducer Mechanism: G Protein–Coupled Receptors
G Protein–Coupled Receptors (GPCRs) are membrane-bound receptors that transiently associate with heterotrimeric G proteins and induce an appropriate response to various stimuli. GPCRs regulate critical physiological pathways and are excellent drug targets for treating diseases such as diabetes, cancer, obesity, depression, or Alzheimer's. Nearly 35% of approved drugs implement their therapeutic effects by selectively interacting with specific GPCRs.
GPCRs are also called heptahelical, 7TM, or...
GPCRs are also called heptahelical, 7TM, or...