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SM-IMPORT SIGNED

Substrate import at work: single-molecule studies of ABC transporters

Total Cost €

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EC-Contrib. €

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Partnership

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Project "SM-IMPORT" data sheet

The following table provides information about the project.

Coordinator
LUDWIG-MAXIMILIANS-UNIVERSITAET MUENCHEN 

Organization address
address: GESCHWISTER SCHOLL PLATZ 1
city: MUENCHEN
postcode: 80539
website: www.uni-muenchen.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]
 Total cost 1˙500˙000 €
 EC max contribution 1˙500˙000 € (100%)
 Programme 1. H2020-EU.1.1. (EXCELLENT SCIENCE - European Research Council (ERC))
 Code Call ERC-2014-STG
 Funding Scheme ERC-STG
 Starting year 2015
 Duration (year-month-day) from 2015-09-01   to  2020-12-31

 Partnership

Take a look of project's partnership.

# participants  country  role  EC contrib. [€] 
1    LUDWIG-MAXIMILIANS-UNIVERSITAET MUENCHEN DE (MUENCHEN) coordinator 500˙000.00
2    RIJKSUNIVERSITEIT GRONINGEN NL (GRONINGEN) participant 1˙000˙000.00

Map

 Project objective

ABC (ATP Binding Cassette) transporters represent the most abundant and diverse family of transport proteins known that play crucial roles in numerous cellular processes. Despite their importance, all proposed molecular models for transport are based on indirect evidence due to the inability of classical biophysical and biochemical techniques to directly visualize dynamic structural changes. To solve this problem, I suggest a novel approach that can decipher the molecular mechanisms of transport, with the ultimate goal to use this knowledge against pathogenic bacteria, for treatment of ABC-related diseases or multi-drug resistance of cancer cells. I propose to use single-molecule fluorescence microscopy for the study of conformational states of an ABC model system in vitro, and thus to observe directly how elementary transport steps are coordinated. This will open up a virtually unexplored biophysical research area and provide a detailed understanding of the molecular mechanisms of ABC transporters. The key questions of this proposal are:

Aim 1: What is the mechanism of substrate binding in ABC transporters? The conformational equilibrium (open vs. closed state) of ABC-associated substrate-binding proteins will be studied to understand the molecular mechanism of binding, i.e., induced fit vs. conformational selection.

Aim 2: What are relevant conformational states and changes for substrate translocation? The time- and length-scales of conformational changes in transmembrane and nucleotide binding domains as well as interactions with other domains will be characterized using the osmoregulator OpuA as a model system.

Aim 3: How are substrate binding, energy utilization and translocation coordinated in ABC transporters? Finally, a complete model of transport will be developed to decipher the coordination of transport events, i.e., how substrate binding and ATP-hydrolysis are coupled and transferred into conformational changes that drive substrate transport.

 Publications

year authors and title journal last update
List of publications.
2018 Eitan Lerner, Thorben Cordes, Antonino Ingargiola, Yazan Alhadid, SangYoon Chung, Xavier Michalet, Shimon Weiss
Toward dynamic structural biology: Two decades of single-molecule Förster resonance energy transfer
published pages: eaan1133, ISSN: 0036-8075, DOI: 10.1126/science.aan1133
Science 359/6373 2020-03-20
2019 Jochem H. Smit, Jasper H. M. van der Velde, Jingyi Huang, Vanessa Trauschke, Sarah S. Henrikus, Si Chen, Nikolaos Eleftheriadis, Eliza M. Warszawik, Andreas Herrmann, Thorben Cordes
On the impact of competing intra- and intermolecular triplet-state quenching on photobleaching and photoswitching kinetics of organic fluorophores
published pages: 3721-3733, ISSN: 1463-9076, DOI: 10.1039/c8cp05063e
Physical Chemistry Chemical Physics 21/7 2020-03-20
2018 Florence Husada, Kiran Bountra, Konstantinos Tassis, Marijn de Boer, Maria Romano, Sylvie Rebuffat, Konstantinos Beis, Thorben Cordes
Conformational dynamics of the ABC transporter McjD seen by single‐molecule FRET
published pages: e100056, ISSN: 0261-4189, DOI: 10.15252/embj.2018100056
The EMBO Journal 37/21 2020-03-20
2018 Rebecca Mächtel, Christian Gebhardt, Thorben Cordes
Konformationsbewegungen von aktiven Membrantransportern
published pages: 495-497, ISSN: 0947-0867, DOI: 10.1007/s12268-018-0945-2
BIOspektrum 24/5 2020-03-20
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 2020-03-20
2019 Jasper H M van der Velde, Jochem H Smit, Elke Hebisch, Michiel Punter, Thorben Cordes
Self-healing dyes for super-resolution fluorescence microscopy
published pages: 34001, ISSN: 0022-3727, DOI: 10.1088/1361-6463/aae752
Journal of Physics D: Applied Physics 52/3 2020-03-20
2016 Nicole C. Robb, Aartjan J. W. te Velthuis, Ralph Wieneke, Robert Tampé, Thorben Cordes, Ervin Fodor, Achillefs N. Kapanidis
Single-molecule FRET reveals the pre-initiation and initiation conformations of influenza virus promoter RNA
published pages: gkw884, ISSN: 0305-1048, DOI: 10.1093/nar/gkw884
Nucleic Acids Research (2016) 44 (21): 10304-10315 2020-03-20
2016 Jasper H. M. van der Velde, Jens Oelerich, Jingyi Huang, Jochem H. Smit, Atieh Aminian Jazi, Silvia Galiani, Kirill Kolmakov, Giorgos Guoridis, Christian Eggeling, Andreas Herrmann, Gerard Roelfes, Thorben Cordes
A simple and versatile design concept for fluorophore derivatives with intramolecular photostabilization
published pages: 10144, ISSN: 2041-1723, DOI: 10.1038/ncomms10144
Nature Communications 7 2020-03-20
2016 Eitan Lerner, Evelyn Ploetz, Johannes Hohlbein, Thorben Cordes, Shimon Weiss
A Quantitative Theoretical Framework For Protein-Induced Fluorescence Enhancement–Förster-Type Resonance Energy Transfer (PIFE-FRET)
published pages: 6401-6410, ISSN: 1520-6106, DOI: 10.1021/acs.jpcb.6b03692
The Journal of Physical Chemistry B 120/26 2020-03-20
2016 Evelyn Ploetz, Eitan Lerner, Florence Husada, Martin Roelfs, SangYoon Chung, Johannes Hohlbein, Shimon Weiss, Thorben Cordes
Förster resonance energy transfer and protein-induced fluorescence enhancement as synergetic multi-scale molecular rulers
published pages: , ISSN: 2045-2322, DOI: 10.1038/srep33257
Scientific Reports 6/1 2020-03-20
2017 Atieh Aminian Jazi, Evelyn Ploetz, Muhamad Arizki, Balasubramaniam Dhandayuthapani, Izabela Waclawska, Reinhard Krämer, Christine Ziegler, Thorben Cordes
Caging and Photoactivation in Single-Molecule Förster Resonance Energy Transfer Experiments
published pages: 2031-2041, ISSN: 0006-2960, DOI: 10.1021/acs.biochem.6b00916
Biochemistry 56/14 2020-03-20

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