The ability of GPCRs to assemble into multimeric complexes is one of the most recently studied and discussed topics for many reasons, including the possibility that GPCR assemblies show a distinct pharmacological profile offering an innovative avenue for the drug synthesis. In addition, the possible differential coupling of monomeric versus multimeric GPCRs to G proteins and other downstream partners, as well as the signaling, the regulation through desensitization and internalization, and the subcellular localization can well represent additional factors that contribute to GPCR-mediated physiopathological states. The standard biochemical techniques used to identify GPCR interactions, such as coimmunoprecipitation, have obvious limitations owing to the use of nonphysiological buffers and detergents that disrupt the natural cell environment and biological interactions and preclude the analysis of subcellular localization and compartmentalization. In the past decade, new biophysical proximity assays based on the resonance energy transfer (RET) between two chromophores allow the study of dimerization in intact living cells, thus proving more information on GPCR physiological roles. In this chapter, we detail the application of two RET techniques based on fluorescence (FRET) and bioluminescence (BRET) to the study of GPCR dimerization and describe the results that can be obtained

Busnelli, M., Mauri, M., Parenti, M., Chini, B. (2013). Analysis of GPCR dimerization using acceptor photobleaching resonance energy transfer techniques.. In P.M. Conn (a cura di), G Protein Coupled Receptors: Trafficking and Oligomerization (pp. 311-327). Academic Press [10.1016/B978-0-12-391862-8.00017-X].

Analysis of GPCR dimerization using acceptor photobleaching resonance energy transfer techniques.

MAURI, MARIO;PARENTI, MARCO DOMENICO;
2013

Abstract

The ability of GPCRs to assemble into multimeric complexes is one of the most recently studied and discussed topics for many reasons, including the possibility that GPCR assemblies show a distinct pharmacological profile offering an innovative avenue for the drug synthesis. In addition, the possible differential coupling of monomeric versus multimeric GPCRs to G proteins and other downstream partners, as well as the signaling, the regulation through desensitization and internalization, and the subcellular localization can well represent additional factors that contribute to GPCR-mediated physiopathological states. The standard biochemical techniques used to identify GPCR interactions, such as coimmunoprecipitation, have obvious limitations owing to the use of nonphysiological buffers and detergents that disrupt the natural cell environment and biological interactions and preclude the analysis of subcellular localization and compartmentalization. In the past decade, new biophysical proximity assays based on the resonance energy transfer (RET) between two chromophores allow the study of dimerization in intact living cells, thus proving more information on GPCR physiological roles. In this chapter, we detail the application of two RET techniques based on fluorescence (FRET) and bioluminescence (BRET) to the study of GPCR dimerization and describe the results that can be obtained
Capitolo o saggio
BRET; FRET; Receptor dimerization; Thromboxane receptor; Oxytocin receptor
English
G Protein Coupled Receptors: Trafficking and Oligomerization
Conn, PM
2013
978-0-12-391862-8
521
Academic Press
311
327
Busnelli, M., Mauri, M., Parenti, M., Chini, B. (2013). Analysis of GPCR dimerization using acceptor photobleaching resonance energy transfer techniques.. In P.M. Conn (a cura di), G Protein Coupled Receptors: Trafficking and Oligomerization (pp. 311-327). Academic Press [10.1016/B978-0-12-391862-8.00017-X].
none
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/44396
Citazioni
  • Scopus 11
  • ???jsp.display-item.citation.isi??? 9
Social impact