Related Experiment Video

Updated: Jan 29, 2026

Identification of EGFR and RAS Inhibitors using Caenorhabditis elegans
08:12

Identification of EGFR and RAS Inhibitors using Caenorhabditis elegans

Published on: October 5, 2020

3.4K

Bioluminescence Resonance Energy Transfer 2 (BRET2)-Based RAS Biosensors to Characterize RAS Inhibitors

Nicolas Bery1, Terence H Rabbitts1

  • 1Weatherall Institute of Molecular Medicine, MRC Molecular Haematology Unit, University of Oxford, John Radcliffe Hospital, Headington, Oxford, United Kingdom.

Current Protocols in Cell Biology
|February 16, 2019
PubMed

Insights

This study introduces a novel Bioluminescence Resonance Energy Transfer 2 (BRET2)-based RAS biosensor. This method effectively detects and characterizes RAS protein-protein interaction inhibition for potential cancer therapeutics.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • Protein-protein interactions (PPIs) are vital for cellular functions and implicated in diseases like cancer.
  • Targeting PPIs for therapeutic intervention remains challenging, particularly RAS-effector interactions in cancers.
  • Existing methods for studying PPIs have limitations in characterizing inhibitors.

Purpose of the Study:

  • To present a protocol for a novel Bioluminescence Resonance Energy Transfer 2 (BRET2)-based RAS biosensor.
  • To demonstrate the utility of this biosensor for detecting and characterizing RAS PPI inhibition.
  • To provide a versatile platform applicable to other small GTPases and PPIs.

Main Methods:

  • Development and application of BRET2-based biosensors specific for RAS proteins.
  • Utilizing the BRET2 system to monitor RAS protein-protein interactions in real-time.
  • Characterization of inhibition of RAS PPIs by both small molecules and macromolecules.

Main Results:

  • The BRET2-based RAS biosensor successfully detected and characterized RAS PPI inhibition.
  • The protocol provides a sensitive and specific method for evaluating potential PPI inhibitors.
  • The approach is adaptable for studying other small GTPase interactions.

Conclusions:

  • BRET2-based RAS biosensors offer a powerful tool for studying RAS PPIs and drug discovery.
  • This method facilitates the development of novel therapeutics targeting challenging PPIs.
  • The protocol's adaptability makes it valuable for broader PPI research.
Keywords:
BRETRAS family GTPasesbiosensorsprotein-protein interaction inhibition

More Related Videos

Author Spotlight: Integrating BRET-Based Assays and Rare Mutation Analysis to Decipher RAF Kinase Regulation in Live Cells06:44

Author Spotlight: Integrating BRET-Based Assays and Rare Mutation Analysis to Decipher RAF Kinase Regulation in Live Cells

Published on: March 1, 2024

1.7K
Visualizing Cellular Gibberellin Levels Using the nlsGPS1 Förster Resonance Energy Transfer (FRET) Biosensor08:53

Visualizing Cellular Gibberellin Levels Using the nlsGPS1 Förster Resonance Energy Transfer (FRET) Biosensor

Published on: January 12, 2019

11.4K

Related Concept Videos

The Ras Gene02:38

The Ras Gene

The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
Ras is a...
7.3K
Small GTPases - Ras and Rho01:24

Small GTPases - Ras and Rho

Ras and Rho are small monomeric GTPases that act downstream of receptor tyrosine kinase (RTK) and regulate various cellular processes. These GTPases switch between active and inactive states by binding to guanine nucleotides.
Three regulatory proteins control their activity:
5.3K
Resonance02:52

Resonance

The Lewis structure of a nitrite anion (NO2−) may actually be drawn in two different ways, distinguished by the locations of the N-O and N=O bonds.
65.1K
Energy Transfer in Chemical Reactions01:16

Energy Transfer in Chemical Reactions

Chemical reactions require sufficient energy to cause the matter to collide with enough precision and force that old chemical bonds can be broken and new ones formed. In general, kinetic energy is the form of energy powering any type of matter in motion. Imagine a person building a brick wall. The energy it takes to lift and place one brick on top of another is the kinetic energy—the energy matter possesses because of its motion. Once the wall is in place, it stores potential energy.
11.0K
Eukaryotic Transcription Inhibitors01:52

Eukaryotic Transcription Inhibitors

Certain biochemical processes, such as embryonic development and cell growth regulation, depend on the repression of specific genes. DNA binding proteins known as eukaryotic transcription inhibitors regulate the repression of gene expression in eukaryotes. The presence of these inhibitors at the required location and time in the cell is triggered by the presence of hormones and additional signals from other cells.
Eukaryotic transcription inhibitors usually contain two distinct domains, a...
11.0K
Ionization Energy03:12

Ionization Energy

The amount of energy required to remove the most loosely bound electron from a gaseous atom in its ground state is called its first ionization energy (IE1). The first ionization energy for an element, X, is the energy required to form a cation with 1+ charge:
43.3K