Abstract
Background and PurposeThe mechanosensitive Piezo1 channel has important roles in vascular physiology and disease. Yoda1 is a small‐molecule agonist, but the pharmacology of these channels is otherwise limited.Experimental ApproachYoda1 analogues were generated by synthetic chemistry. Intracellular Ca2+ and Tl+ measurements were made in HEK 293 or CHO cell lines overexpressing channel subunits and in HUVECs, which natively express Piezo1. Isometric tension recordings were made from rings of mouse thoracic aorta.Key ResultsModification of the pyrazine ring of Yoda1 yielded an analogue, which lacked agonist activity but reversibly antagonized Yoda1. The analogue is referred to as Dooku1. Dooku1 inhibited 2 μM Yoda1‐induced Ca2+‐entry with IC50s of 1.3 μM (HEK 293 cells) and 1.5 μM (HUVECs) yet failed to inhibit constitutive Piezo1 channel activity. It had no effect on endogenous ATP‐evoked Ca2+ elevation or store‐operated Ca2+ entry in HEK 293 cells or Ca2+ entry through TRPV4 or TRPC4 channels overexpressed in CHO and HEK 293 cells. Yoda1 caused dose‐dependent relaxation of aortic rings, which was mediated by an endothelium‐ and NO‐dependent mechanism and which was antagonized by Dooku1 and analogues of Dooku1.Conclusion and ImplicationsChemical antagonism of Yoda1‐evoked Piezo1 channel activity is possible, and the existence of a specific chemical interaction site is suggested with distinct binding and efficacy domains.
Highlights
Piezo[1] protein is important for mechanical force sensing and its transduction in higher organisms (Coste et al, 2010; Ranade et al, 2015; Wu et al, 2016)
This study has provided insight into the structure–activity relationships for Piezo[1] channel activation by Yoda[1] with the goal of generating new tools for investigating Piezo[1] channel function
We have identified and named Dooku[1], an inhibitor of Yoda1-induced Piezo[1] channel activity that strongly inhibits Yoda1-induced
Summary
Piezo[1] protein is important for mechanical force sensing and its transduction in higher organisms (Coste et al, 2010; Ranade et al, 2015; Wu et al, 2016). It assembles as a trimer with a propeller-like structure around a central ion pore, which is permeable to the cations Na+, K+ and Ca2+ (Coste et al, 2012; 2015; Ge et al, 2015; Guo and MacKinnon, 2017; Saotome et al, 2017; Wu et al, 2017; Zhao et al, 2018). Mechanical forces that include membrane tension and laminar flow are able to activate the channel (Coste et al, 2010; Li et al, 2014; Lewis and Grandl, 2015; Syeda et al, 2016). Isometric tension recordings were made from rings of mouse thoracic aorta
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