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 Receptors01:15

G Protein-coupled Receptors

11.0K
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...
11.0K
Transducer Mechanism: G Protein–Coupled Receptors01:30

Transducer Mechanism: G Protein–Coupled Receptors

1.8K
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,...
1.8K
GPCRs Regulate Adenylyl Cylase Activity01:09

GPCRs Regulate Adenylyl Cylase Activity

5.1K
Some GPCRs transmit signals through adenylyl cyclase (AC), a transmembrane enzyme. AC helps synthesize second messenger cyclic adenosine monophosphate (cAMP). AC catalyzes cyclization reaction and converts ATP to cAMP by releasing a pyrophosphate. The pyrophosphate is further hydrolyzed to phosphate by the enzyme pyrophosphatase, which drives cAMP synthesis to completion. However, cAMP is rapidly degraded to 5′ AMP by the enzymes phosphodiesterase (PDE), preventing overstimulation of...
5.1K
GPCR Desensitization01:12

GPCR Desensitization

5.7K
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...
5.7K
G-protein Coupled Receptors01:21

G-protein Coupled Receptors

114.2K
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.
114.2K
Activation and Inactivation of G Proteins01:22

Activation and Inactivation of G Proteins

6.5K
Heterotrimeric G proteins are guanine nucleotide-binding proteins. As the name suggests, heterotrimeric G proteins are composed of three subunits: alpha, beta, and gamma. They remain GDP-bound or GTP-bound inside the cells and switch between inactive/active states. The Gα subunit possesses the nucleotide-binding pocket that binds guanine nucleotides and switches between GDP or GTP-bound states. In contrast, the Gꞵ and Gγ subunits are always bound together with high...
6.5K

You might also read

Related Articles

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

Sort by
Same author

Tuning the Response of GPCR-Based Yeast Sensors Using Fluorescent Reporters.

ACS synthetic biology·2025
Same author

Cell-Free-Based Thermophilic Biocatalyst for the Synthesis of Amino Acids from One-Carbon Feedstocks.

ACS synthetic biology·2025
Same author

Carbon Negative Synthesis of Amino Acids Using a Cell-Free-Based Biocatalyst.

ACS synthetic biology·2024
Same author

Insight into the Mode of Action of 8-Hydroxyquinoline-Based Blockers on the Histamine Receptor 2.

Biosensors·2023
Same author

Olfactory Receptors as an Emerging Chemical Sensing Scaffold.

Biochemistry·2022
Same author

Discovery of 8-Hydroxyquinoline as a Histamine Receptor 2 Blocker Scaffold.

ACS synthetic biology·2022

Related Experiment Video

Updated: May 23, 2025

G Protein-selective GPCR Conformations Measured Using FRET Sensors in a Live Cell Suspension Fluorometer Assay
09:12

G Protein-selective GPCR Conformations Measured Using FRET Sensors in a Live Cell Suspension Fluorometer Assay

Published on: September 10, 2016

9.6K

Modulating the Properties of GPCR-Based Sensors Via C-Terminus Isoforms.

Paola L Marquez-Gomez1, Sonia R Damiano2, Lily R Torp1

  • 1School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia 30332, United States.

ACS Synthetic Biology
|April 25, 2025
PubMed
Summary

This study introduces GPCR isoforms as a novel method to engineer yeast biosensors. Using serotonin receptor 4 (5-HTR4) isoforms, researchers modulated sensor dynamic and linear ranges for improved chemical detection.

Keywords:
GPCRisoformssensorsserotoninyeast

More Related Videos

A Kinetic Fluorescence-based Ca2+ Mobilization Assay to Identify G Protein-coupled Receptor Agonists, Antagonists, and Allosteric Modulators
07:41

A Kinetic Fluorescence-based Ca2+ Mobilization Assay to Identify G Protein-coupled Receptor Agonists, Antagonists, and Allosteric Modulators

Published on: February 20, 2018

8.8K
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

3.1K

Related Experiment Videos

Last Updated: May 23, 2025

G Protein-selective GPCR Conformations Measured Using FRET Sensors in a Live Cell Suspension Fluorometer Assay
09:12

G Protein-selective GPCR Conformations Measured Using FRET Sensors in a Live Cell Suspension Fluorometer Assay

Published on: September 10, 2016

9.6K
A Kinetic Fluorescence-based Ca2+ Mobilization Assay to Identify G Protein-coupled Receptor Agonists, Antagonists, and Allosteric Modulators
07:41

A Kinetic Fluorescence-based Ca2+ Mobilization Assay to Identify G Protein-coupled Receptor Agonists, Antagonists, and Allosteric Modulators

Published on: February 20, 2018

8.8K
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

3.1K

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Chemical Biology

Background:

  • G-protein coupled receptors (GPCRs) are crucial for chemical biosensing, detecting diverse molecules like hormones and neurotransmitters.
  • GPCR-based biosensors in yeast offer rapid engineering potential via coupling to the yeast mating pathway, yielding fluorescent or luminescent signals.
  • Modulating GPCR sensor properties, such as dynamic and linear ranges, typically requires complex engineering of yeast cellular machinery.

Purpose of the Study:

  • To investigate the utility of GPCR C-terminal isoforms for modulating the properties of engineered chemical biosensors.
  • To demonstrate a novel strategy for rapidly tuning the dynamic and linear ranges of GPCR-based biosensors in yeast.

Main Methods:

  • Leveraged nine naturally occurring serotonin receptor 4 (5-HTR4) C-terminus isoforms to construct serotonin biosensors.
  • Integrated these isoforms into yeast mating pathways to create single and double integrated sensor versions.
  • Characterized the dynamic and linear ranges of the resulting biosensors upon serotonin activation.

Main Results:

  • Serotonin sensors constructed with 5-HTR4 C-terminal isoforms exhibited dynamic ranges from 2- to 8.5-fold (single integrated) and 3.4- to 62.7-fold (double integrated).
  • Linear ranges extended up to five orders of magnitude (10^-8 to 10^-3 M serotonin).
  • Observed differential sensor performance based on the activating chemical, suggesting potential isoform-specific activation in vivo.

Conclusions:

  • GPCR C-terminal isoforms represent a new and effective strategy for rapidly modulating the dynamic and linear ranges of GPCR-based biosensors.
  • This approach simplifies the engineering of yeast biosensors with tailored detection capabilities.
  • The observed differential activation hints at the physiological relevance of 5-HTR4 C-terminal isoforms in the human body.