title: Differential efficacy of GoSlo-SR compounds on BKα and BKαγ1-4channels.
abstract: Large conductance, voltage and Ca2+activated K+channels (BK channels) are abundantly expressed throughout the body and are important regulators of smooth muscle tone and neuronal excitability. Their dysfunction is implicated in various diseases including overactive bladder, hypertension and erectile dysfunction. Therefore, BK channel openers bear significant therapeutic potential to treat the above diseases. GoSlo-SR compounds were designed to be potent and efficacious BK channel openers. Although their structural activity relationships, activation in both BKα and BKαβ channels and the hypothetical mode of action of these compounds has been studied in detail in recent years, their effectiveness to open the BKαγ channels still remains unexplored. In this study, we have examined the efficacy of 3 closely related GoSlo-SR openers, GoSlo-SR-5-6 (SR-5-6), GoSlo-SR-5-44 (SR-5-44) and GoSlo-SR-5-130 (SR-5-130) using inside out patches on BKα channels coexpressed with 4 different LRRC (γ1-4) subunits in HEK293 cells. Our data suggests that the activation effects due to SR-5-6 were not significantly affected in the presence of γ1-4subunits. Interestingly, the effects of more efficacious BK channel opener SR-5-44 were altered by different γ subunits. In cells expressing BKα channels, the shift in V1/2(ΔV1/2) induced by SR-5-44 (3 μM) was -76 ± 3 mV, whereas it was significantly reduced by ∼70 % in BKαγ1channels (ΔV1/2= -23 ± 3, p < 0.001, ANOVA). In BKαγ2channels the ΔV1/2was -36 ± 1 mV, which was less than that observed in BKαγ3and BKαγ4channels where the ΔV1/2was -47 ± 5 mV, and -82 ± 5 mV, respectively. Additionally, the excitatory effects of a 'β specific' BK channel opener, SR-5-130 were only partially restored in the patches containing BKαγ1-4channels. Together this data highlights that subtle modifications in GoSlo-SR structures alter their effectiveness on BK channels with accessory γ subunits and this study might provide a scaffold for the development of more tissue specific BK channel openers.
authors.affiliation: a The Smooth Muscle Research Center, Dundalk Institute of Technology , Dundalk , Ireland.
authors.forename: Aravind S
authors.initials: AS
authors.lastname: Kshatri
authors.affiliation: b Department of Anesthesiology and Preoperative Medicine , The University of Texas MD Anderson Cancer Center , Houston , TX , USA.
authors.forename: Qin
authors.initials: Q
authors.lastname: Li
authors.affiliation: b Department of Anesthesiology and Preoperative Medicine , The University of Texas MD Anderson Cancer Center , Houston , TX , USA.
authors.forename: Jiusheng
authors.initials: J
authors.lastname: Yan
authors.affiliation: a The Smooth Muscle Research Center, Dundalk Institute of Technology , Dundalk , Ireland.|c Ion Channel Biotechnology Center, Dundalk Institute of Technology , Dundalk , Ireland.
authors.forename: Roddy J
authors.initials: RJ
authors.lastname: Large
authors.affiliation: a The Smooth Muscle Research Center, Dundalk Institute of Technology , Dundalk , Ireland.|c Ion Channel Biotechnology Center, Dundalk Institute of Technology , Dundalk , Ireland.
authors.forename: Gerard P
authors.initials: GP
authors.lastname: Sergeant
authors.affiliation: a The Smooth Muscle Research Center, Dundalk Institute of Technology , Dundalk , Ireland.|c Ion Channel Biotechnology Center, Dundalk Institute of Technology , Dundalk , Ireland.
authors.forename: Noel G
authors.initials: NG
authors.lastname: McHale
authors.affiliation: a The Smooth Muscle Research Center, Dundalk Institute of Technology , Dundalk , Ireland.|c Ion Channel Biotechnology Center, Dundalk Institute of Technology , Dundalk , Ireland.
authors.forename: Keith D
authors.initials: KD
authors.lastname: Thornbury
authors.affiliation: a The Smooth Muscle Research Center, Dundalk Institute of Technology , Dundalk , Ireland.|c Ion Channel Biotechnology Center, Dundalk Institute of Technology , Dundalk , Ireland.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Hollywood
mesh_terms: D000880:Anthraquinones / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000235:genetics / Q000502:physiology*; D021122:Protein Subunits / Q000235:genetics / Q000502:physiology; D013451:Sulfonic Acids / Q000494:pharmacology*
chemical_list: D000880:Anthraquinones; C000607770:GoSlo-SR-5-130; C000623365:GoSlo-SR-5-44; C000607769:GoSlo-SR-5-6; D051036:Large-Conductance Calcium-Activated Potassium Channels; D021122:Protein Subunits; D013451:Sulfonic Acids
keywords: BK channels; GoSlo-SR openers; activation; drug binding; electrophysiology; γ subunits