Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

G-protein Coupled Receptors01:21

G-protein Coupled Receptors

134.2K
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.
134.2K
Labeling DNA Probes03:31

Labeling DNA Probes

9.7K
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.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
9.7K
G Protein-coupled Receptors01:15

G Protein-coupled Receptors

19.6K
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.
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...
19.6K
Transducer Mechanism: G Protein–Coupled Receptors01:30

Transducer Mechanism: G Protein–Coupled Receptors

7.7K
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,...
7.7K
Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

8.7K
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
8.7K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Single-molecule dynamics reveal ATP binding alone powers substrate translocation by an ABC transporter.

Nature communications·2026
Same author

Architectural principles of transporter-chaperone coupling within the native MHC I peptide-loading complex.

Science advances·2026
Same author

Human Substance P Interactions with G Protein-Coupled Receptor NK1R Observed by NMR in Solution.

Journal of the American Chemical Society·2025
Same author

Meet the authors: Canyong Guo and Kurt Wüthrich.

Cell chemical biology·2025
Same author

Structural basis of adenosine 2A receptor-balanced signaling activation relies on allosterically mediated structural dynamics.

Cell chemical biology·2025
Same author

Rimota-Gd: Paramagnetic Probe for In Vivo MRI Studies of the Cannabinoid 1 Receptor Distribution in the Mouse Brain.

ACS chemical neuroscience·2024

Related Experiment Video

Updated: Mar 30, 2026

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors
16:16

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Published on: September 13, 2013

15.9K

In-Membrane Chemical Modification (IMCM) for Site-Specific Chromophore Labeling of GPCRs.

Lukas Sušac1, Casey O'Connor1,2, Raymond C Stevens2

  • 1Department of Integrative Structural and Computational Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037 (USA).

Angewandte Chemie (International Ed. in English)
|November 8, 2015
PubMed
Summary

We developed in-membrane chemical modification (IMCM) to selectively label G-protein-coupled receptors (GPCRs) with minimal genetic changes. This technique enables precise chromophore introduction for advanced spectroscopy studies of GPCRs.

Keywords:
G-protein-coupled receptorsNMR spectroscopyfluorescence spectroscopymembrane proteinssite-specific labeling

More Related Videos

Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
14:02

Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells

Published on: April 9, 2018

9.2K
Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET
10:59

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET

Published on: August 17, 2022

3.9K

Related Experiment Videos

Last Updated: Mar 30, 2026

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors
16:16

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Published on: September 13, 2013

15.9K
Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
14:02

Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells

Published on: April 9, 2018

9.2K
Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET
10:59

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET

Published on: August 17, 2022

3.9K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Spectroscopy

Background:

  • G-protein-coupled receptors (GPCRs) are crucial cell surface proteins involved in numerous physiological processes.
  • Studying GPCR structure and function often requires specific labeling for spectroscopic techniques.
  • Existing labeling methods can be limited by accessibility and the need for extensive genetic manipulation.

Purpose of the Study:

  • To introduce a novel method, in-membrane chemical modification (IMCM), for selective labeling of GPCRs.
  • To enable site-specific introduction of chromophores for spectroscopic analysis of GPCRs.
  • To demonstrate the utility of IMCM in studying specific GPCRs like the κ-opioid receptor (KOR) and A2A adenosine receptor (A2A AR).

Main Methods:

  • Utilizing the membrane environment to protect most cysteines, allowing selective modification of exposed cysteines.
  • Applying IMCM for site-specific chromophore attachment to intracellular cysteines in GPCRs.
  • Employing NMR and fluorescence spectroscopy to analyze labeled GPCRs.

Main Results:

  • Successfully demonstrated selective chromophore labeling of intracellular cysteines in GPCRs using IMCM.
  • Showcased the application of IMCM in the human KOR and A2A AR, enabling detailed spectroscopic studies.
  • Validated IMCM as a method requiring minimal mutagenesis for effective labeling.

Conclusions:

  • In-membrane chemical modification (IMCM) provides a powerful tool for selective GPCR labeling.
  • IMCM facilitates site-specific introduction of spectroscopic probes with minimal genetic alteration.
  • This method is broadly applicable to in vitro GPCR studies and holds promise for in-cell spectroscopy.