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PROSINT SIGNED

Multi-protein interaction kinetics by single molecule methods

Total Cost €

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EC-Contrib. €

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Partnership

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Project "PROSINT" data sheet

The following table provides information about the project.

Coordinator
ALBERT-LUDWIGS-UNIVERSITAET FREIBURG 

Organization address
address: FAHNENBERGPLATZ
city: FREIBURG
postcode: 79098
website: www.uni-freiburg.de

contact info
title: n.a.
name: n.a.
surname: n.a.
function: n.a.
email: n.a.
telephone: n.a.
fax: n.a.

 Coordinator Country Germany [DE]
 Project website http://www.singlemolecule.uni-freiburg.de
 Total cost 1˙882˙500 €
 EC max contribution 1˙882˙500 € (100%)
 Programme 1. H2020-EU.1.1. (EXCELLENT SCIENCE - European Research Council (ERC))
 Code Call ERC-2015-CoG
 Funding Scheme ERC-COG
 Starting year 2016
 Duration (year-month-day) from 2016-04-01   to  2021-03-31

 Partnership

Take a look of project's partnership.

# participants  country  role  EC contrib. [€] 
1    ALBERT-LUDWIGS-UNIVERSITAET FREIBURG DE (FREIBURG) coordinator 1˙882˙500.00

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 Project 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.

 Publications

year authors and title journal last update
List of publications.
2018 Björn Hellenkamp, Sonja Schmid, Olga Doroshenko, Oleg Opanasyuk, Ralf Kühnemuth, Soheila Rezaei Adariani, Benjamin Ambrose, Mikayel Aznauryan, Anders Barth, Victoria Birkedal, Mark E. Bowen, Hongtao Chen, Thorben Cordes, Tobias Eilert, Carel Fijen, Christian Gebhardt, Markus Götz, Giorgos Gouridis, Enrico Gratton, Taekjip Ha, Pengyu Hao, Christian A. Hanke, Andreas Hartmann, Jelle Hendrix, Lasse L. Hildebrandt, Verena Hirschfeld, Johannes Hohlbein, Boyang Hua, Christian G. Hübner, Eleni Kallis, Achillefs N. Kapanidis, Jae-Yeol Kim, Georg Krainer, Don C. Lamb, Nam Ki Lee, Edward A. Lemke, Brié Levesque, Marcia Levitus, James J. McCann, Nikolaus Naredi-Rainer, Daniel Nettels, Thuy Ngo, Ruoyi Qiu, Nicole C. Robb, Carlheinz Röcker, Hugo Sanabria, Michael Schlierf, Tim Schröder, Benjamin Schuler, Henning Seidel, Lisa Streit, Johann Thurn, Philip Tinnefeld, Swati Tyagi, Niels Vandenberk, Andrés Manuel Vera, Keith R. Weninger, Bettina Wünsch, Inna S. Yanez-Orozco, Jens Michaelis, Claus A. M. Seidel, Timothy D. Craggs, Thorsten Hugel
Precision and accuracy of single-molecule FRET measurements—a multi-laboratory benchmark study
published pages: 669-676, ISSN: 1548-7091, DOI: 10.1038/s41592-018-0085-0
Nature Methods 15/9 2019-06-18
2017 Philipp Wortmann, Markus Götz, Thorsten Hugel
Cooperative Nucleotide Binding in Hsp90 and Its Regulation by Aha1
published pages: 1711-1718, ISSN: 0006-3495, DOI: 10.1016/j.bpj.2017.08.032
Biophysical Journal 113/8 2019-06-18
2016 Sonja Schmid, Markus Götz, Thorsten Hugel
Single-Molecule Analysis beyond Dwell Times: Demonstration and Assessment in and out of Equilibrium
published pages: 1375-1384, ISSN: 0006-3495, DOI: 10.1016/j.bpj.2016.08.023
Biophysical Journal 111/7 2019-06-18
2016 Björn Hellenkamp, Philipp Wortmann, Florian Kandzia, Martin Zacharias, Thorsten Hugel
Multidomain structure and correlated dynamics determined by self-consistent FRET networks
published pages: 174-180, ISSN: 1548-7091, DOI: 10.1038/nmeth.4081
Nature Methods 14/2 2019-06-18
2018 Sonja Schmid, Markus Götz, Thorsten Hugel
Effects of Inhibitors on Hsp90′s Conformational Dynamics, Cochaperone and Client Interactions
published pages: 1716-1721, ISSN: 1439-4235, DOI: 10.1002/cphc.201800342
ChemPhysChem 19/14 2019-05-08
2018 Björn Hellenkamp, Sonja Schmid, Olga Doroshenko, Oleg Opanasyuk, Ralf Kühnemuth, Soheila Rezaei Adariani, Benjamin Ambrose, Mikayel Aznauryan, Anders Barth, Victoria Birkedal, Mark E. Bowen, Hongtao Chen, Thorben Cordes, Tobias Eilert, Carel Fijen, Christian Gebhardt, Markus Götz, Giorgos Gouridis, Enrico Gratton, Taekjip Ha, Pengyu Hao, Christian A. Hanke, Andreas Hartmann, Jelle Hendrix, Lasse L. Hildebrandt, Verena Hirschfeld, Johannes Hohlbein, Boyang Hua, Christian G. Hübner, Eleni Kallis, Achillefs N. Kapanidis, Jae-Yeol Kim, Georg Krainer, Don C. Lamb, Nam Ki Lee, Edward A. Lemke, Brié Levesque, Marcia Levitus, James J. McCann, Nikolaus Naredi-Rainer, Daniel Nettels, Thuy Ngo, Ruoyi Qiu, Nicole C. Robb, Carlheinz Röcker, Hugo Sanabria, Michael Schlierf, Tim Schröder, Benjamin Schuler, Henning Seidel, Lisa Streit, Johann Thurn, Philip Tinnefeld, Swati Tyagi, Niels Vandenberk, Andrés Manuel Vera, Keith R. Weninger, Bettina Wünsch, Inna S. Yanez-Orozco, Jens Michaelis, Claus A. M. Seidel, Timothy D. Craggs, Thorsten Hugel
Precision and accuracy of single-molecule FRET measurements—a multi-laboratory benchmark study
published pages: 669-676, ISSN: 1548-7091, DOI: 10.1038/s41592-018-0085-0
Nature Methods 15/9 2019-05-08
2018 Weixiang Ye, Markus Götz, Sirin Celiksoy, Laura Tüting, Christoph Ratzke, Janak Prasad, Julia Ricken, Seraphine V. Wegner, Rubén Ahijado-Guzmán, Thorsten Hugel, Carsten Sönnichsen
Conformational Dynamics of a Single Protein Monitored for 24 h at Video Rate
published pages: 6633-6637, ISSN: 1530-6984, DOI: 10.1021/acs.nanolett.8b03342
Nano Letters 18/10 2019-05-08
2018 Katarzyna M. Tych, Markus Jahn, Florian Gegenfurtner, Vera K. Hechtl, Johannes Buchner, Thorsten Hugel, Matthias Rief
Nucleotide-Dependent Dimer Association and Dissociation of the Chaperone Hsp90
published pages: , ISSN: 1520-6106, DOI: 10.1021/acs.jpcb.8b07301
The Journal of Physical Chemistry B 2019-05-08
2018 Sonja Schmid, Thorsten Hugel
Efficient use of single molecule time traces to resolve kinetic rates, models and uncertainties
published pages: 123312, ISSN: 0021-9606, DOI: 10.1063/1.5006604
The Journal of Chemical Physics 148/12 2019-05-08
2018 Björn Hellenkamp, Johann Thurn, Martina Stadlmeier, Thorsten Hugel
Kinetics of Transient Protein Complexes Determined via Diffusion-Independent Microfluidic Mixing and Fluorescence Stoichiometry
published pages: , ISSN: 1520-6106, DOI: 10.1021/acs.jpcb.8b07437
The Journal of Physical Chemistry B 2019-05-08

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