Thursday, September 11, 2014

Applications: BRET beta-Arrestin Interaction Assay

Bioluminescence resonance energy transfer, or BRET for short, has been used for many years now by scientists seeking to monitor molecular interactions such as protein-protein interactions.

General bioluminescence reaction of coelenterazine
by Yikrazuul
The original method used Renilla luciferase (Rluc) and yellow flourescent protein (YFP). In the example of protein-protein interactions one protein is labeled with Rluc and another with YFP. When the proteins associate the Rluc and YFP are brought into proximity, such that when the Rluc substrate coelenterazine is present Rluc produces light whose energy can be transfered to the YFP acceptor.

The BRET method has seen several modifications over the years including NanoBRET which was discussed in a BMG LABTECH webinar last year. However, a recent JBC paper employs the classic BRET approach to analyze the interaction of the long-chain fatty acid receptor, FFA4, with beta-arrestin in order to study which ligands bind to FFA4.

This analysis employed the PHERAstar FS from BMG LABTECH. When the FS is equipped with BRET 1 optic module it can perform simultaneous dual emission detection and measure the luminescent emission at 530 and 490 nm produced by YFP and Rluc respectively.

For more information on how BMG LABTECH equipment can help you measure BRET and many other applications please visit our website: http://www.bmglabtech.com/

Original article citation: B.D. Hudson, et al. "The Molecular Basis of Ligand Interaction at Free Fatty Acid Receptor 4 (FFA4/GPR120)" J Biol Chem. 2014 289(29): 20345–20358

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