TY - JOUR
T1 - Two transmembrane Cys residues are involved in 5-HT4 receptor dimerization
AU - Berthouze, Magali
AU - Rivail, Lucie
AU - Lucas, Alexandre
AU - Ayoub, Mohammed A.
AU - Russo, Olivier
AU - Sicsic, Sames
AU - Fischmeister, Rodolphe
AU - Berque-Bestel, Isabelle
AU - Jockers, Ralf
AU - Lezoualc'h, Frank
N1 - Funding Information:
Magali Berthouze was supported by a pre-doctoral grant from the Ministère de la Recherche et de l’Enseignement Supérieur and the Fondation pour la Recherche Médicale. We thank Monique Gastineau, Mireille Giner and Jean-Luc Guillaume for their technical assistance.
PY - 2007/5/11
Y1 - 2007/5/11
N2 - The 5-HT4 receptor (5-HT4R) belongs to the G-protein-coupled receptor (GPCR) family and is of considerable interest for the development of new drugs to treat gastrointestinal diseases and memory disorders. The 5-HT4R exists as a constitutive dimer but its molecular determinants are still unknown. Using co-immunoprecipitation and Bioluminescence Resonance Energy Transfer (BRET) techniques, we show here that 5-HT4R homodimerization but not 5-HT4R-β2 adrenergic receptor (β2AR) heterodimerization is largely decreased under reducing conditions suggesting the participation of disulfide bonds in 5-HT4R dimerization. Molecular modeling and protein docking experiments identified four cysteine (Cys) residues potentially involved in the dimer interface through intramolecular or intermolecular disulfide bonds. We show that disulfide bridges between Cys112 and Cys145 located within TM3 and TM4, respectively, are of critical importance for 5-HT4R dimer formation. Our data suggest that two disulfide bridges between two transmembrane Cys residues are involved in the dimerization interface of a GPCR.
AB - The 5-HT4 receptor (5-HT4R) belongs to the G-protein-coupled receptor (GPCR) family and is of considerable interest for the development of new drugs to treat gastrointestinal diseases and memory disorders. The 5-HT4R exists as a constitutive dimer but its molecular determinants are still unknown. Using co-immunoprecipitation and Bioluminescence Resonance Energy Transfer (BRET) techniques, we show here that 5-HT4R homodimerization but not 5-HT4R-β2 adrenergic receptor (β2AR) heterodimerization is largely decreased under reducing conditions suggesting the participation of disulfide bonds in 5-HT4R dimerization. Molecular modeling and protein docking experiments identified four cysteine (Cys) residues potentially involved in the dimer interface through intramolecular or intermolecular disulfide bonds. We show that disulfide bridges between Cys112 and Cys145 located within TM3 and TM4, respectively, are of critical importance for 5-HT4R dimer formation. Our data suggest that two disulfide bridges between two transmembrane Cys residues are involved in the dimerization interface of a GPCR.
KW - BRET
KW - Dimer
KW - Disulfide bridge
KW - G-protein-coupled receptor
KW - Molecular modeling
KW - Serotonin
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U2 - 10.1016/j.bbrc.2007.03.030
DO - 10.1016/j.bbrc.2007.03.030
M3 - Article
C2 - 17379184
AN - SCOPUS:34047107637
SN - 0006-291X
VL - 356
SP - 642
EP - 647
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -