Cel Single-molecule enzymology offers new possibilities to dissect catalytic reactions that were previously unapproachable using biochemistry techniques conducted in the bulk. In particular, recent discoveries conducted at the single molecule level, such as the unanticipated force-mediated protein degradation pathway in the proteasome, highlight the close relation between mechanical forces and proteolysis in vivo. While much has been discovered about protein enzymology in the recent decades, the question of how mechanical force affects enzymatic catalysis remains vastly elusive. The main goal of this proposal is to understand the mechanobiology of proteolysis at the single molecule level. We will use the newly developed force-clamp spectroscopy technique, together with molecular biology engineering techniques and bioinformatics structural analysis to elucidate the molecular mechanisms that underlie protease catalysis under mechanical force. Successful enzymatic activity relies on the enzyme:substrate (E:S) assembly. Upon mechanical unfolding, proteins unveil their buried substrate sites, also called cryptic sites, thus favoring the formation of the E:S complex and ultimately permitting the subsequent chemical reaction. A key feature of recent mechano-chemistry experiments at the single bond level is that the rate at which the reduction of a protein disulfide bond occurs in the presence of a nucleophile is exponentially dependent on the stretching force. Hence, it is tempting to speculate that, in the case of an enzymatic reaction, the catalytic rate will be also force-dependent. We anticipate that the curved geometry of the bound substrate inhibits the E:S assembly at high-forces, implying a novel mechano-specificity character of proteases. Within a multidisciplinary approach, here we propose a series of innovative experiments to directly probe the effect of force on the kinetics of protease hydrolysis. Dziedzina nauki natural sciencesphysical sciencesopticsmicroscopynatural sciencesphysical sciencesclassical mechanicsstatistical mechanicsnatural scienceschemical sciencescatalysisnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymesnatural sciencesbiological sciencesmolecular biology Program(-y) H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions Main Programme H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility Temat(-y) MSCA-IF-2014-EF - Marie Skłodowska-Curie Individual Fellowships (IF-EF) Zaproszenie do składania wniosków H2020-MSCA-IF-2014 Zobacz inne projekty w ramach tego zaproszenia System finansowania MSCA-IF-EF-ST - Standard EF Koordynator KING'S COLLEGE LONDON Wkład UE netto € 195 454,80 Adres STRAND WC2R 2LS London Zjednoczone Królestwo Zobacz na mapie Region London Inner London — West Westminster Rodzaj działalności Higher or Secondary Education Establishments Linki Kontakt z organizacją Opens in new window Strona internetowa Opens in new window Uczestnictwo w unijnych programach w zakresie badań i innowacji Opens in new window sieć współpracy HORIZON Opens in new window Koszt całkowity € 195 454,80