Negative cooperativity across β1-adrenoceptor homodimers provides insights into the nature of the secondary low-affinity CGP 12177 β1-adrenoceptor binding conformation

Karolina Gherbi, Lauren T. May, Jillian G Baker, Stephen J Briddon, Stephen J Hill

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At the β1-adrenoceptor, CGP 12177 potently antagonizes agonist responses at the primary highaffinity catecholamine conformation while also exerting agonist effects of its own through a secondary low-affinity conformation. A recent mutagenesis study identified transmembrane region (TM)4 of the β1-adrenoceptor as key for this low-affinity conformation. Others suggested that TM4 has a role in β1-adrenoceptor oligomerization. Here, assessment of the dissociation rate of a fluorescent analog of CGP 12177 [bordifluoropyrromethane-tetramethylrhodamine-(±)CGP 12177 (BODIPY-TMR-CGP)] at the human β1-adrenoceptor expressed in Chinese hamster ovary cells revealed negative cooperative interactions between 2 distinct β1-adrenoceptor conformations. The dissociation rate of 3 nM BODIPY-TMR-CGP was 0.09 ± 0.01 min-1 in the absence of competitor ligands, and this was enhanced 2.2- and 2.1-fold in the presence of 1 μM CGP 12177 and 1 μM propranolol, respectively. These effects on the BODIPY-TMR-CGP dissociation rate were markedly enhanced in β1-adrenoceptor homodimers constrainedbybimolecular fluorescencecomplementation(9.8-and9.9-foldfor1μM CGP 12177 and 1 μM propranolol, respectively) and abolished in β1-adrenoceptors containing TM4 mutations vital for the second conformation pharmacology. This study suggests that negative cooperativity across a β1-adrenoceptor homodimer may be responsible for generating the low-affinity pharmacology of the secondary β1-adrenoceptor conformation.

Original languageEnglish
Pages (from-to)2859-2871
Number of pages13
JournalThe FASEB Journal
Issue number7
Publication statusPublished - 1 Jul 2015
Externally publishedYes


  • Allosterism
  • Dissociation
  • GPCR
  • Receptor dimerization

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