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Related Concept Videos

G-protein Coupled Receptors01:21

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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.
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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
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G Protein-Coupled Receptors or GPCRs are membrane-bound receptors that transiently associate with heterotrimeric G proteins and induce an appropriate response to sensory stimuli such as light, odors, hormones, cytokines, or neurotransmitters.
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Related Experiment Video

Updated: Sep 17, 2025

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors
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Probing Antibody Binding Sites on G Protein-Coupled Receptors Using Genetically Encoded Photo-Activatable

Victoria R Saca1,2, Jordan M Mattheisen1,2, Thomas Huber1

  • 1Laboratory of Chemical Biology and Signal Transduction, The Rockefeller University, New York, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 2, 2025
PubMed
Summary

Researchers developed a new method to map antibody binding sites on G protein-coupled receptors (GPCRs). This technique uses genetic code expansion and photo-activatable amino acids to pinpoint interactions, aiding drug development.

Keywords:
Amber codonAntibodyEpitope mapG protein-coupled receptorPhoto-activatable cross-linkers

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • G protein-coupled receptors (GPCRs) are crucial drug targets.
  • Monoclonal antibodies (mAbs) are vital tools for studying GPCRs.
  • Precisely mapping mAb epitopes on GPCRs is essential for understanding interactions.

Purpose of the Study:

  • To present a novel methodology for mapping the epitopes of monoclonal antibodies (mAbs) that bind to G protein-coupled receptors (GPCRs).
  • To enable the precise localization of antibody binding sites on receptor surfaces.
  • To provide a versatile method applicable to various cell-surface proteins.

Main Methods:

  • Utilized genetic code expansion to incorporate photo-activatable non-canonical amino acids (ncAAs) like azF or BzF into GPCRs.
  • Employed ultraviolet (UV) irradiation to induce cross-linking between engineered GPCRs and cognate mAbs.
  • Mapped interaction sites by identifying cross-linked ncAA residues and comparing them with mutation data causing binding loss.

Main Results:

  • Successfully generated surface binding maps of mAb epitopes on GPCRs.
  • Demonstrated that the method can elucidate discontinuous epitopes.
  • Showcased the applicability of the technique beyond GPCRs to other cell-surface proteins.

Conclusions:

  • The described methodology offers a precise approach to map antibody epitopes on GPCRs and other cell-surface proteins.
  • This technique enhances the understanding of antibody-target interactions, facilitating antibody-based therapeutics and diagnostics.
  • The precision of the epitope map is influenced by the number of mutants and the availability of structural models.