Objective Objective of this proposal is to obtain a real time picture of how components of a multi-protein complex interact to perform complex regulated tasks, in particular to see how a protein system might be more than the sum of its components. In living organisms many proteins work in complexes to form multicomponent protein machines and to regulate cellular processes. The function of such multicomponent machines is usually addressed by dividing them into a collection of two state systems at equilibrium. Many molecular machines work in large complexes with multiple states out of equilibrium by utilizing the energy of ATP hydrolysis. In this proposal the real time kinetics of multi-protein interactions in and out of equilibrium will be investigated using single molecule methods. The succession of association and dissociation steps as well as large conformational changes within the proteins will be monitored simultaneously with multicolour single molecule FRET in real time. In order to observe at the same time the folding state of proteins, a combination of optical tweezers and single molecule FRET will be developed. Finally, a combination of microfluidics for fast mixing and single pair FRET will be developed to investigate interactions of low affinity. A large part of the experiments will focus on the example of the heat shock protein Hsp90 system, which consists of co-chaperones, clients and nucleotides. Altogether, these interdisciplinary experiments will yield unprecedented information on multi-component interactions in equilibrium and out of equilibrium on timescales from sub milliseconds to hours. I am confident that these studies will have an impact on the understanding of the Hsp90 machinery as well as general principles of multi-component protein systems, which is the basis for understanding cellular processes. Fields of science natural sciencescomputer and information sciencesdata sciencenatural sciencesphysical sciencesclassical mechanicsfluid mechanicsmicrofluidicsnatural sciencescomputer and information sciencessoftwarenatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural sciencesbiological sciencesgeneticsnucleotides Programme(s) H2020-EU.1.1. - EXCELLENT SCIENCE - European Research Council (ERC) Main Programme Topic(s) ERC-CoG-2015 - ERC Consolidator Grant Call for proposal ERC-2015-CoG See other projects for this call Funding Scheme ERC-COG - Consolidator Grant Host institution ALBERT-LUDWIGS-UNIVERSITAET FREIBURG Net EU contribution € 1 882 500,00 Address FAHNENBERGPLATZ 79098 Freiburg Germany See on map Region Baden-Württemberg Freiburg Freiburg im Breisgau, Stadtkreis Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 1 882 500,00 Beneficiaries (1) Sort alphabetically Sort by Net EU contribution Expand all Collapse all ALBERT-LUDWIGS-UNIVERSITAET FREIBURG Germany Net EU contribution € 1 882 500,00 Address FAHNENBERGPLATZ 79098 Freiburg See on map Region Baden-Württemberg Freiburg Freiburg im Breisgau, Stadtkreis Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 1 882 500,00