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SM-IMPORT SIGNED

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

Total Cost €

EC-Contrib. €

Partnership

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Outcomes and
results

SM-IMPORT project word cloud

Explore the words cloud of the SM-IMPORT project. It provides you a very rough idea of what is the project "SM-IMPORT" about.

atp indirect model fluorescence interactions dynamic microscopy unexplored biophysical substrate drive complete visualize single time vitro drug virtually coupled abundant suggest models events decipher binding fit vs closed conformational molecule coordinated transport nucleotide diverse transmembrane scales understand despite inability cancer mechanisms cellular area translocation treatment questions transporters diseases length observe structural cassette hydrolysis opua transferred proteins family equilibrium elementary roles play domains biochemical resistance abc pathogenic cells bacteria energy mechanism ultimate molecular techniques osmoregulator

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 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.	DE (MUENCHEN)	coordinator	500˙000.00
2	RIJKSUNIVERSITEIT GRONINGEN RIJKSUNIVERSITEIT GRONINGEN Organization address address: Broerstraat 5 city: GRONINGEN postcode: 9712CP website: www.rug.nl contact info title: n.a. name: n.a. surname: n.a. function: n.a. email: n.a. telephone: n.a. fax: n.a.	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

List of publications.
year	authors and title	journal	last update
2018	Eitan Lerner, Thorben Cordes, Antonino Ingargiola, Yazan Alhadid, SangYoon Chung, Xavier Michalet, Shimon Weiss Toward dynamic structural biology: Two decades of single-molecule FÃ¶rster 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 singleâ€molecule FRET published pages: e100056, ISSN: 0261-4189, DOI: 10.15252/embj.2018100056	The EMBO Journal 37/21	2020-03-20
2018	Rebecca MÃ¤chtel, 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	BjÃ¶rn Hellenkamp, Sonja Schmid, Olga Doroshenko, Oleg Opanasyuk, Ralf KÃ¼hnemuth, 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 GÃ¶tz, 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. HÃ¼bner, Eleni Kallis, Achillefs N. Kapanidis, Jae-Yeol Kim, Georg Krainer, Don C. Lamb, Nam Ki Lee, Edward A. Lemke, BriÃ© Levesque, Marcia Levitus, James J. McCann, Nikolaus Naredi-Rainer, Daniel Nettels, Thuy Ngo, Ruoyi Qiu, Nicole C. Robb, Carlheinz RÃ¶cker, Hugo Sanabria, Michael Schlierf, Tim SchrÃ¶der, Benjamin Schuler, Henning Seidel, Lisa Streit, Johann Thurn, Philip Tinnefeld, Swati Tyagi, Niels Vandenberk, AndrÃ©s Manuel Vera, Keith R. Weninger, Bettina WÃ¼nsch, Inna S. Yanez-Orozco, Jens Michaelis, Claus A. M. Seidel, Timothy D. Craggs, Thorsten Hugel Precision and accuracy of single-molecule FRET measurementsâ€”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, AartjanÂ J.Â W. teÂ Velthuis, Ralph Wieneke, Robert TampÃ©, 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 Enhancementâ€“FÃ¶rster-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 FÃ¶rster 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 KrÃ¤mer, Christine Ziegler, Thorben Cordes Caging and Photoactivation in Single-Molecule FÃ¶rster 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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