Arch funds from the Interdisciplinary Center for Clinical Research (Interdisziplinares Zentrum fur Klinische Forschung, IZKF) from the University of Wurzburg, Germany (NU, EW: N-260). NU was supported by the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG: UE 171-5/1)Further informationFundingFunder Interdisziplinares Zentrum fur Klinische Forschung, Universitatsklinikum Wurzburg Deutsche Forschungsgemeinschaft Grant reference 40592-88-9 medchemexpress number N-260 Author Erhard Wischmeyer �� Nurcan Uceyler �� Nurcan UceylerUE 171-5/The funders had no function in study style, data collection and interpretation, or the decision 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 analysis, 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, Data curation, Investigation, Writing–original draft; Erhard Wischmeyer, Information curation, Formal analysis, Funding �� acquisition, Investigation, Methodology, Writing–original draft; Nurcan Uceyler, Conceptualization, Data 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 147116-67-4 site Ethics Animal experimentation: Our study was authorized by the Bavarian State authorities (Regierung von Unterfranken, # 54/12).Choice letter and Author response Choice letter https://doi.org/10.7554/eLife.39300.013 Author response https://doi.org/10.7554/eLife.39300.More filesSupplementary files . Mechanical stimulation of Piezo channels provides rise to a mechanically-activated (MA) existing, which swiftly decays as a consequence of 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 specifically affect this inactivation process, suggesting that the regular timing of MA present decay is vital for animal physiology (Wu et al., 2017a). Furthermore, a prolongation of Piezo2 inactivation in somatosensory neurons of tactile-specialist birds suggests that inactivation is involved within the modulation of complicated behaviors (Schneider et al., 2017; Anderson et al., 2017; Schneider et al., 2014). Inactivation is substantially impacted by the known modulators of Piezo1: Yoda1 and Jedi1/2 (Lacroix et al., 2018; Wang et al., 2018; Evans et al., 2018; Syeda et al., 2015). Yet, 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 exceptional 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). Every blade is composed of 36 transmembrane (TM) segments and is believed to contribute to sensing tension within 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 added.