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Multifunktionelles hochauflösendes Abbilden und Kartieren der Kraftfelder biologischer Membranen und Membranproteine

English title Multifunctional high-resolution imaging and force-field mapping of membrane proteins and biological membranes
Applicant Müller Daniel Jobst
Number 134521
Funding scheme Project funding (Div. I-III)
Research institution Computational Systems Biology Department of Biosystems, D-BSSE ETH Zürich
Institution of higher education ETH Zurich - ETHZ
Main discipline Other disciplines of Physics
Start/End 01.03.2012 - 28.02.2015
Approved amount 378'359.00
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All Disciplines (5)

Discipline
Other disciplines of Physics
Biophysics
Cellular Biology, Cytology
Biochemistry
Molecular Biology

Keywords (5)

Bionanotechnology; Multifunctional High-Resolution Imaging; Structure and Function of Membrane Proteins; Microscopy; Spectroscopy

Lay Summary (English)

Lead
Lay summary

Every biological process depends on the dynamic interplay of molecular interactions. For example biological cells rely on using interactions in a controllable and dynamic manner. Therefore, the timely interaction of correct binding partners is critical in molecular recognition, signal transduction, the assembly of hierarchical and functional structures, cell-cell communication, adhesion, development, and all other cellular activities. Essential for life are cellular membranes through which cells differentiate, communicate, take up and release molecules, and generate energy. Membrane proteins provide cell membranes with unique functions that are precisely controlled and adapted to the cell’s functional need. How interactions guide the dynamic assembly of membrane proteins into functional units and modulate their functional state remains largely unknown. Unfortunately, high-resolution microscopy (≈1 nm) tools that allow the structural characterization of native membrane protein assemblies are rare. Moreover, multifunctional high-resolution microscopic and spectroscopic approaches that enable to structurally observe membrane proteins and to simultaneously quantify their specific and unspecific interactions do not exist. To reveal insight into how biological membrane and membrane proteins are guided by interactions we intend to develop and establish a multifunctional high-resolution microscopic tool that allows to image single native membrane proteins of complex assemblies at sub-nanometer resolution and to quantify the interactions that guide membrane protein assembly and function. We intend to structurally map the interactions of membrane proteins with their environment and the interactions that switch the functional state of membrane proteins. Therefore, we will establish a new atomic force microscopy (AFM) approach that images the surface of biological membranes at a resolution of ≤1 nm and simultaneously maps the interactions of single membrane proteins in three dimensions. First experiments using this microscopic mode revealed a lateral resolution of ≈1.5-2 nm of native membrane proteins and allowed to quantify and to structurally map their interactions. We will further develop and establish this new multifunctional and nanotechnological tool to be able to provide unique structural and quantitative insights into basic biological interactions that direct assembly and function of native proteins of the cell membrane.  

Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
SAS-6 engineering reveals interdependence between cartwheel and microtubules in determining centriole architecture
Hilbert Manuel, Noga Akira, Frey Daniel, Hamel Virginie, Guichard Paul, Kraatz Sebastian, Pfreundschuh Moritz, Hosner Sarah, Flückiger Isabelle, Jaussi Rolf, Wieser Mara M., Thieltges Katharine M., Deupi Xavier, Müller Daniel J., Kammerer Richard A., Gönczy Pierre, Hirono Masafumi, Steinmetz Michel O. (2016), SAS-6 engineering reveals interdependence between cartwheel and microtubules in determining centriole architecture, in Nature Cell Biology, 18, 393-403.
Identifying and quantifying two ligand-binding sites while imaging native human membrane receptors by AFM
Pfreundschuh Moritz, Alsteens David, Wieneke Ralf, Zhang Cheng, Coughlin Shaun R., Tampé Robert, Kobilka Brian K., Müller Daniel J. (2015), Identifying and quantifying two ligand-binding sites while imaging native human membrane receptors by AFM, in Nature Communications , 6, 8857.
Impact of holdase chaperones Skp and SurA on the folding of β-barrel outer membrane proteins
Thoma J., Burmann B.M., Hiller S., Müller D.J. (2015), Impact of holdase chaperones Skp and SurA on the folding of β-barrel outer membrane proteins, in Nature Structural and Molecular Biology , 22, 795-802.
Localizing Chemical Groups while Imaging Single Native Proteins by High-Resolution Atomic Force Microscopy
Pfreundschuh Moritz, Alsteens David, Hilbert Manuel, Steinmetz Michel, Müller Daniel J. (2014), Localizing Chemical Groups while Imaging Single Native Proteins by High-Resolution Atomic Force Microscopy, in Nano Letters, 14(5), 2957-2964.
Multiparametric high-resolution imaging of native proteins by force-distance curve-based AFM
Pfreundschuh Moritz, Martinez-Martin David, Mulvihill Estefania, Wegmann Susanne, Müller Daniel J. (2014), Multiparametric high-resolution imaging of native proteins by force-distance curve-based AFM, in Nature Protocols, 9(5), 1113-1130.
Multi-parametric force mapping of biological systems to molecular resolution
Dufrene Yves. F., Martínez-Martin David, Medalsy Izhar, Alsteens David, Muller Daniel J. (2013), Multi-parametric force mapping of biological systems to molecular resolution, in Nature Methods, 10, 847-854.
Nanomechanical properties of proteins and membranes depend on loading rate and electrostatic interactions
Medalsy Izhar, Muller Daniel J. (2013), Nanomechanical properties of proteins and membranes depend on loading rate and electrostatic interactions, in ACS Nano, 7, 2642-2650.
Quantitative imaging of the electrostatic field and potential generated by a transmembrane protein at sub-nanometer resolution
Pfreundschuh Moritz, Hensen Ulf, Muller Daniel J. (2013), Quantitative imaging of the electrostatic field and potential generated by a transmembrane protein at sub-nanometer resolution, in Nano Letters, 13(11), 5585-5593.
Engineering rotor ring stoichiometries in the ATP synthase
Pogoryelov D, Klyszejko AL, Krasnoselska GO, Heller EM, Leone V, Langer VD, Vonck J, Muller DJ, Faraldo-Gómez JD, Meier T (2012), Engineering rotor ring stoichiometries in the ATP synthase, in Proc Natl Acad Sci U S A, 109(25), E1599-E1608.
Out but not in: the large transmembrane β-barrel protein FhuA unfolds but cannot refold via β-hairpins
Thoma J, Bosshart P, Pfreundschuh M, Müller DJ (2012), Out but not in: the large transmembrane β-barrel protein FhuA unfolds but cannot refold via β-hairpins, in Structure, 20(12), 2185-2190.
The fuzzy coat of pathological human Tau fibrils is a polyelectrolyte polymer brush
Wegmann S., Medalsy I., Mandelkow E., Müller DJ (2012), The fuzzy coat of pathological human Tau fibrils is a polyelectrolyte polymer brush, in Proc. Natl. Acad. Sci. USA, 110(4), E313-321.

Collaboration

Group / person Country
Types of collaboration
Dr. Michel Steinmetz, Paul Scherrer Institut, Villingen Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Dr. Thomas Meier, Max-Planck-Institute of Biophysics, Frankfurt Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof. Dr. Gina Sosinsky, University of California San Diego, School of Medicine United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
Prof. Dr. Brian Kobilka, Stanford University, Dept. Cellular and Molecular Physiology United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof. Dr. E. Mandelkow, MPI für Structural Biology, Hamburg Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof. Dr. Kai Simons, Max-Planck-Institute of Molecular Cell Biology and Genetics, Dresden Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Prof. Dr. Krystof Palczewsky, Cleveland University, School of Medicine, Dept. Pharmacology United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
Membrane Research Forum Talk given at a conference Multifunctional Imaging of native G-Protein Coupled Receptors using FD-based AFM 01.03.2015 Kyoto, Japan Pfreundschuh Moritz;
International Titisee Conference Talk given at a conference Directing the folding pathways of proteins into membranes 08.10.2014 Titisee, Germany Müller Daniel Jobst;
Single Molecule Approaches to Biology, Gordon Research Conference Talk given at a conference Directing the folding of proteins into membranes 13.07.2014 Lucca, Italy Müller Daniel Jobst;
Bi-Annual Meeting of the Spanish Society for Chemical Biology Individual talk Directly observing the unfolding, folding and working of single native membrane proteins at subnanometer resolution 04.03.2014 Bilbao, Spain Müller Daniel Jobst;
58th Annual Meeting of the Biophysical Society Individual talk Quantitative Imaging of the Electrostatic Field and Potential Generated by a Transmembrane Protein Pore at Subnanometer Resolution 07.02.2014 San Francisco, United States of America Pfreundschuh Moritz;
Bi-Annual Meeting of the Spanish Society for Chemical Biology Talk given at a conference Directly observing the unfolding, folding and working of single native membrane proteins at subnanometer resolution 04.02.2014 Bilbo, Spain Müller Daniel Jobst;
Gordon Research Conference (GRC) on Protein Folding Dynamics Talk given at a conference Folding- and unfolding of single transmembrane β-barrel proteins 08.01.2014 Galveston, United States of America Thoma Johannes; Müller Daniel Jobst;
Gordon Research Conference (GRC) on Protein Folding Dynamics Poster Folding- and unfolding- pathways of transmembrane ß-barrel proteins 05.01.2014 Galveston, United States of America Thoma Johannes; Müller Daniel Jobst;
Swiss-Kyoto Symposium for Advanced Bio/Nanotechnology Talk given at a conference Deciphering Molecular Mechanics Guiding Cellular Processes using Nanoscopic Assays 21.11.2013 Zurich, Switzerland Müller Daniel Jobst;
UK SPM 2013 Meeting Individual talk Deciphering Molecular Mechanics Guiding Cellular Processes using Nanoscopic Assays 26.06.2013 Leeds, Great Britain and Northern Ireland Müller Daniel Jobst;
Global Challenges - Opportunities for Nanotechnology Poster High Resolution Force Volume AFM Detects Electrostatics of the Transmembrane Porin OmpF 15.04.2013 Venedig, Italy Pfreundschuh Moritz;
ACRITAS Meeting Talk given at a conference High-resolution Imaging of Native Ion Channels by FD-based AFM 27.02.2013 Dublin, Ireland Müller Daniel Jobst;
AFM in Biology Workshop Individual talk Guiding receptors to control neuronal recognition and guide neuronal outgrowth 05.11.2012 Kanazawa, Japan, Japan Müller Daniel Jobst;
SPM on Soft and Polymeric Materials Individual talk Multifunctional High-Resolution AFM of Native Proteins In Vivo and In Vitro 24.09.2012 Reduc Abbey, Netherlands., Netherlands Müller Daniel Jobst;
17th European Bioenergetics Conference Individual talk Multi-parametric 3D force nanoscopy of cellular machines at molecular resolution 17.09.2012 Freiburg, Germany, Germany Müller Daniel Jobst;
International Conference on Nanosciene and Technology Individual talk Multi-parametric 3D force nanoscopy of cellular machines at molecular resolution 24.07.2012 Paris, France Müller Daniel Jobst;
Spanish Biophysical Society Meeting Individual talk Molecular Mechanics Guiding Cellular Processes 03.07.2012 Barcelona, Spain, Spain Müller Daniel Jobst;
Swiss Nanoconvention Individual talk Bionanotools to image, quantify and manipulate the cellular machinery at molecular resolution 23.05.2012 Lausanne, Switzerland Müller Daniel Jobst;


Self-organised

Title Date Place

Awards

Title Year
European Microscopy Society Outstanding Paper Award 2015 of the Paper D. Alsteens et al. Nature Methods (2015) 12, 845-851. 2015

Associated projects

Number Title Start Funding scheme
160199 Gleichzeitiges hochauflösendes Abbilden von Membranproteinen und dreidimensionales Kartieren der Wechselwirkungskräfte und Energielandschaften mit Liganden 01.03.2016 Project funding (Div. I-III)
138063 Systematische Charakterisierung der Kommunikation von Zelladhäsionsrezeptoren mittels Einzelzellkraftspektroskopie 01.09.2012 Project funding (Div. I-III)

Abstract

We intend to develop and establish a multifunctional high-resolution tool that allows imaging of single native membrane proteins of complex assemblies at sub-nanometer resolution and quantifying of interactions that guide membrane protein assembly and function. We intend to structurally map the interactions of membrane proteins with their environment and the interactions inside membrane proteins that switch their functional state. To this end, we will establish and develop a new atomic force microscopy (AFM) approach that images the surface of biological membranes at a resolution of =1 nm and simultaneously maps the interactions of single membrane proteins in three dimensions. Preliminary experiments reveal a lateral resolution of ˜1.5-2 nm of native membrane proteins using this new mode. We will further develop this method to be able to image native membrane receptors at a resolution of =1 nm and at the same time to detect how a ligand finds its way at far distance from the membrane surface (˜100 nm) to bind specifically to the receptor’s binding pocket. Furthermore, the assay will quantify and map interactions established upon ligand-binding and the interactions that switch the protein’s functional state. Characterizing how different ligands establish characteristic interactions will teach us how membrane receptors are functionally modulated by interactions. Moreover, the method will be also applicable to quantify and to structurally map the physico-chemical properties of the native membrane receptor surface and the interaction of any given molecule with the membrane receptor surface. With this new multifunctional and nanotechnological tool we will be able to provide unique structural and quantitative insights into basic biological interactions that direct assembly and function of native proteins of the cell membrane.
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