Arch funds with the Interdisciplinary Center for Clinical Analysis (Interdisziplinares Zentrum fur Klinische Forschung, IZKF) in the University of Wurzburg, Germany (NU, EW: N-260). NU was supported by the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG: UE 171-5/1)Added informationFundingFunder Interdisziplinares Zentrum fur Klinische Forschung, Universitatsklinikum Wurzburg Deutsche Forschungsgemeinschaft Grant reference number N-260 6729-55-1 web Author Erhard Wischmeyer �� Acetylcholine (iodide) In Vivo Nurcan Uceyler �� Nurcan UceylerUE 171-5/The funders had no function in study style, data collection and interpretation, or the choice to submit the function for publication. Author contributions Lukas Hofmann, Formal analysis, Investigation, Methodology, Writing–original draft; Dorothea Hose, Anne Grie ammer, Robert Blum, Formal evaluation, Investigation, Writing–review and editing; Frank Doring, Investigation, Writing–review and editing; Sulayman Dib-Hajj, Stephen Waxman, Methodology, Writing–review and editing; Claudia Sommer, Conceptualization, Information curation, Investigation, Writing–original draft; Erhard Wischmeyer, Information curation, Formal evaluation, Funding �� acquisition, Investigation, Methodology, Writing–original draft; Nurcan Uceyler, Conceptualization, Information curation, Formal evaluation, Supervision, Funding acquisition, Investigation, Methodology, Writing–original draft, Project administration Author ORCIDs Lukas Hofmann http://orcid.org/0000-0002-8397-1819 Sulayman Dib-Hajj http://orcid.org/0000-0002-4137-1655 �� Nurcan Uceyler http://orcid.org/0000-0001-6973-6428 Ethics Animal experimentation: Our study was approved by the Bavarian State authorities (Regierung von Unterfranken, # 54/12).Selection letter and Author response Decision letter https://doi.org/10.7554/eLife.39300.013 Author response https://doi.org/10.7554/eLife.39300.Extra filesSupplementary files . Mechanical stimulation of Piezo channels gives rise to a mechanically-activated (MA) present, which speedily decays resulting from quick inactivation (Lewis et al., 2017; Gottlieb et al., 2012). Disease-linkedZheng et al. eLife 2019;8:e44003. DOI: https://doi.org/10.7554/eLife.1 ofResearch articleStructural Biology and Molecular Biophysicsmutations in Piezo1 and Piezo2 particularly have an effect on this inactivation method, suggesting that the normal timing of MA present decay is vital for animal physiology (Wu et al., 2017a). In addition, a prolongation of Piezo2 inactivation in somatosensory neurons of tactile-specialist birds suggests that inactivation is involved within the modulation of complex behaviors (Schneider et al., 2017; Anderson et al., 2017; Schneider et al., 2014). Inactivation is significantly impacted by the identified modulators of Piezo1: Yoda1 and Jedi1/2 (Lacroix et al., 2018; Wang et al., 2018; Evans et al., 2018; Syeda et al., 2015). But, in spite of its significance for channel function, physiology and pathophysiology, the mechanism of Piezo inactivation remains unknown. Functional Piezo channels are homo-trimers that adopt a unique propeller-like architecture comprising a central C-terminal ion-conducting pore and 3 peripheral N-terminal blades (Figure 1A) (Guo and MacKinnon, 2017; Saotome et al., 2018; Zhao et al., 2018). Each and every blade is composed of 36 transmembrane (TM) segments and is believed to contribute to sensing tension in the membrane (Guo and MacKinnon, 2017; Haselwandter and MacKinnon, 2018). The pore area, which consists of an outer pore helix (OH), an inner pore helix (IH), an further.