Biophysics and Molecular Materials
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benoit_martin

Dr. Martin Benoit

Responsibilities

advanced practical courses & assistant researcher

topics:
cell mechanics
membrane forces
cell adhesion forces
methods:
force measurements (AFM )
planar patch-clamp

Contact

Lehrstuhl für Angewandte Physik
und Center for NanoScience
Ludwig-Maximilians-Universität
Amalienstr. 54
80799 München

Room: 006
Phone: +49 (0) 89/ 2180-3133
Fax: +49 (0) 89/ 2180-3150

Office hours:
tuesday 14.00 - 16.00 p.m.

Further Information

Publications:

loef2017jcp_figweb_kleinA Biophysical View on von Willebrand Factor Activation

Achim Löf, Jochen P. Müller, and Maria A. Brehm, Journal of Cellular Physiology,
doi: 10.1002/jcp.25887, March 2017

The process of hemostatic plug formation at sites of vascular injury crucially relies on the large multimeric plasma glycoprotein von Willebrand factor (VWF) and its ability to recruit platelets to the damaged vessel wall via interaction of its A1 domain with platelet GPIbα. Under normal blood flow conditions, VWF multimers exhibit a very low binding affinity for platelets. Only when subjected to increased hydrodynamic forces, which primarily occur in connection with vascular injury, VWF can efficiently bind to platelets. This force-regulation of VWF’s hemostatic activity is not only highly intriguing from a biophysical perspective, but also of eminent physiological importance...  more  PDF

 

loef_aibr017_figwebsiteBiophysical approaches promote advances in the understanding of von Willebrand factor processing and function

Löf A., Müller J.P., Benoit M., Brehm M.A.,
Advances in Biological Regulation 2017, 63:81-91
doi: 10.1016/j.jbior.2016.09.010.

The large multimeric plasma glycoprotein von Willebrand factor (VWF) is essential for primary hemostasis by recruiting platelets to sites of vascular injury. VWF multimers respond to elevated hydrodynamic forces by elongation, thereby increasing their adhesiveness to platelets. Thus, the activation of VWF is force-induced, as is its inactivation. Due to these attributes, VWF is a highly interesting system from a biophysical point of view, and is well suited for investigation using biophysical approaches. Here, we give an overview on recent studies that predominantly employed biophysical methods to gain novel insights into multiple aspects of VWF: Electron microscopy was used to shed light on the domain structure of VWF and the mechanism of VWF secretion...  more  PDF

 

 


muellerloefpH-Dependent Interactions in Dimers Govern the Mechanics and Structure of von Willebrand Factor

Jochen P. Müller, Achim Löf, Salomé Mielke, Tobias Obser, Linda K. Bruetzel, Willem Vanderlinden, Jan Lipfert, Reinhard Schneppenheim, and Martin Benoit, Biophysical Journal,  Volume 111, Issue 2, Pages 312–322, doi:10.1016/j.bpj.2016.06.022, 26 July 2016,

Von Willebrand factor (VWF) is a multimeric plasma glycoprotein that is activated for hemostasis by increased hydrodynamic forces at sites of vascular injury. Here, we present data from atomic force microscopy-based single-molecule force measurements, atomic force microscopy imaging, and small-angle x-ray scattering to show that the structure and mechanics of VWF are governed by multiple pH-dependent interactions with opposite trends within dimeric subunits. In particular, the recently discovered strong intermonomer interaction, which induces a firmly closed conformation of dimers and crucially involves the D4 domain...  more  PDF

 

 

2015-pnas_mueller

Force sensing by the vascular protein von Willebrand factor is tuned by a strong intermonomer interaction

Jochen P. Müller, Salomé Mielke, Achim Löf, Tobias Obser, Christof Beer, Linda K. Bruetzel, Diana A. Pippig, Willem Vanderlinden, Jan Lipfert, Reinhard Schneppenheim, Martin Benoit
PNAS, doi: 10.1073/pnas.1516214113, January 2016

The large plasma glycoprotein von Willebrand factor (VWF) senses hydrodynamic forces in the bloodstream and responds to elevated forces with abrupt elongation, thereby increasing its adhesiveness to platelets and collagen. Remarkably, forces on VWF are elevated at sites of vascular injury, where VWF’s hemostatic potential is important to mediate platelet aggregation and to recruit platelets to the subendothelial layer... more   PDF 

 

 

ediz1Decoding Cytoskeleton-Anchored and Non-Anchored Receptors from Single-Cell Adhesion Force Data

Ediz Sariisik, Cvetan Popov, Jochen P. Müller, Denitsa Docheva, Hauke Clausen-Schaumann and Martin Benoit
Biophys J. 2015 Oct 6;109(7):1330-3. doi: 10.1016/j.bpj.2015.07.048

Complementary to parameters established for cell-adhesion force curve analysis, we evaluated the slope before a force step together with the distance from the surface at which the step occurs and visualized the result in a two-dimensional density plot. This new tool allows detachment steps of long membrane tethers to be distinguished from shorter jumplike force steps.... more   PDF

 


CnidoinA fast recoiling silk-like elastomer facilitates nanosecond nematocyst discharge.

Beckmann A, Xiao S, Müller JP, Mercadante D, Nüchter T, Kröger N, Langhojer F, Petrich W, Holstein T, Benoit M, Gräter F, Özbek S.
BMC Biol. 2015 Jan 16;13:3. doi: 10.1186/s12915-014-0113-1

The discharge of the Cnidarian stinging organelle, the nematocyst, is one of the fastest processes in biology and involves volume changes of the highly pressurised (150 bar) capsule of up to 50%. Hitherto, the molecular basis for the unusual biomechanical properties of nematocysts has been elusive, as their structure was mainly defined as a stress-resistant collagenous matrix. Here, we characterise Cnidoin, a novel elastic protein... more   PDF

 

exponentialExponential size distribution of von Willebrand factor.

Lippok S, Obser T, Müller JP, Stierle VK, Benoit M, Budde U, Schneppenheim R, Rädler JO.
Biophys J. 2013 Sep 3;105(5):1208-16. doi: 10.1016/j.bpj.2013.07.037.

Von Willebrand Factor (VWF) is a multimeric protein crucial for hemostasis. Under shear flow, it acts as a mechanosensor responding with a size-dependent globule-stretch transition to increasing shear rates. Here, we quantify for the first time, to our knowledge, the size distribution of recombinant VWF and VWF-eGFP using a multilateral approach that involves quantitative gel analysis, fluorescence correlation spectroscopy, and total internal reflection fluorescence microscopy... more   PDF

 

 

apracticalA practical guide to quantify cell adhesion using single-cell force spectroscopy.

Friedrichs J, Legate KR, Schubert R, Bharadwaj M, Werner C, Müller DJ, Benoit M.
Methods. 2013 Apr 1;60(2):169-78. doi: 10.1016/j.ymeth.2013.01.006. Epub 2013 Feb 8.

Quantitative analysis of cellular interactions with the extracellular environment is necessary to gain an understanding of how cells regulate adhesion in the development and maintenance of multicellular organisms, and how changes in cell adhesion contribute to diseases. We provide a practical guide to quantify the adhesive strength of living animal cells to various substrates using atomic force microscopy (AFM)-based single-cell force spectroscopy (SCFS)... more   PDF

 

 

eye2013Vital dyes increase the rigidity of the internal limiting membrane

Haritoglou C, Mauell S, Benoit M, Schumann RG, Henrich PB, Wolf A and Kampik A
Eye advance online publication, 16 August 2013; doi:10.1038/eye.2013.178

To assess the stiffness of the natural human internal limiting membrane (ILM) and evaluate potential changes of the mechanical properties following staining with brilliant blue (BB) and indocyanine green (ICG).
Methods Unstained ILM specimens were obtained during ophthalmic surgical procedures. After removal, the specimens were dissected into five parts. .... more   PDF

 

 

cataractIncrease in lens capsule stiffness caused by vital dyes

Christos Haritoglou, MD, Stephan Mauell, MSc, Ricarda G. Schumann, MD, Paul B. Henrich, MD, Armin Wolf, MD, Marcus Kernt, MD, Martin Benoit, PhD
J Cataract Refract Surg 2013; 39:1749–1752 Q 2013 ASCRS and ESCRS, doi:10.1016/j.jcrs.2013.02.057

Cataract surgery was among the first ophthalmic surgical procedures in which dyes were introduced to assist the surgeon in more challenging cases, such as eyes with mature cataract. In mature white cataracts, a controlled capsulorhexis of the anterior capsule is often difficult to performdue to absence of the red fundus reflex. .... more   PDF


 

probingProbing the interaction forces of prostate cancer cells with collagen I and bone marrow derived stem cells on the single cell level.

Sarıısık E, Docheva D, Padula D, Popov C, Opfer J, Schieker M, Clausen-Schaumann H, Benoit M.
Published: March 5, 2013 DOI: 10.1371/journal.pone.0057706, PubMed PMID: 23472100

Adhesion of metastasizing prostate carcinoma cells was quantified for two carcinoma model cell lines LNCaP (lymph node-specific) and PC3 (bone marrow-specific). By time-lapse microscopy and force spectroscopy we found PC3 cells to preferentially adhere to bone marrow-derived mesenchymal stem cells (SCP1 cell line)... more   PDF

 

 

measuringcellMeasuring cell adhesion forces: theory and principles.

Benoit M, Selhuber-Unkel C.  
Methods Mol Biol;736:355-77. PubMed PMID: 21660737.

Cell adhesion is an essential prerequisite for survival, communication, and navigation of cells in organisms. It is maintained by the organized binding of molecules from the cell membrane to the extracellular space. This chapter focuses on direct measurements of cellular binding strength at the level of single adhesion molecules... more   PDF

 

forcespectroscForce Spectroscopy on Cells.

M. Benoit, 2011
Chapter 9, in: handbook of nanophysics 7; 9:1-29; ISBN 978-1-4200-7546-5

... Physicists say: “a cell…” while biologists say, “we took an endothelial cell from
the upper third of the dorsal endometrium in the early S2-phase of a 12 days old male…” For a cell-adhesion measurement with molecular resolution, the biophysicist has a dilemma. The information from the individual adhesion molecule is embedded in the concert of all the participating molecules of the cell including
the membrane and the cytoskeleton... PDF

 

2009 Cuerrier CM, Benoit M, Guillemette G, Gobeil F Jr, Grandbois M.
Real-time monitoring of angiotensin II-induced contractile response and cytoskeleton remodeling in individual cells by atomic force microscopy.
Pflugers Arch;457(6):1361-72.
PubMed PMID: 18953565.   PDF

2008 Schmitz J, Benoit M and Gottschalk KE,
The viscoelasticity of membrane tethers and its importance for cell adhesion.
Biophys J, 95(3):1448–1459
PubMed PMID: 18456832   PDF

2008 Pamir E, George M, Fertig N, Benoit M.
Planar patch-clamp force microscopy on living cells.
Ultramicroscopy;108(6):552-7.
PubMed PMID: 17933465.   PDF

2007 Lugmaier RA, Schedin S, Kühner F, Benoit M.
Dynamic restacking of Escherichia coli P-pili.
Eur Biophys J;37(2):111-20.
PubMed PMID: 17554533.   PDF

2006 Muñoz Javier A, Kreft O, Piera Alberola A, Kirchner C, Zebli B, Susha AS, Horn E, Kempter S, Skirtach AG, Rogach AL, Rädler J, Sukhorukov GB, Benoit M, Parak WJ.
Combined atomic force microscopy and optical microscopy measurements as a method to investigate particle uptake by cells.
Small;2(3):394-400.
PubMed PMID: 17193058.   PDF

2006 Nüchter T, Benoit M, Engel U, Ozbek S, Holstein TW
Nanosecond-scale kinetics of nematocyst discharge.
Curr Biol;16(9):R316-8
PMID:16682335   PDF

2005 Lugmaier R, Hugel T, Benoit M and Gaub HE,
Phase contrast and DIC illumination for AFM hybrids.
Ultramicroscopy, 104(3-4):255–260
PubMed PMID: 15961230.   PDF

2005 Alon R, Feigelson SW, Manevich E, Rose DM, Schmitz J, Overby DR, Winter E, Grabovsky V, Shinder V, Matthews BD, Sokolovsky-Eisenberg M, Ingber DE, Benoit M, Ginsberg MH.J
Alpha4beta1-dependent adhesion strengthening under mechanical strain is regulated by paxillin association with the alpha4-cytoplasmic domain.
Cell Biol;171(6):1073-84.
PubMed PMID: 16365170.   PDF

2005 Neuert G, Kufer S, Benoit M and Gaub HE,
Modular multichannel surface plasmon spectrometer.
Review Of Scientific Instruments, 76(5):054303.   PDF

2004 Blank K, Lankenau A, Mai T, Schiffmann S, Gilbert I, Hirler S, Albrecht C, Benoit M, Gaub HE and Clausen-Schaumann H,
Double-chip protein arrays: force-based multiplex sandwich immunoassays with increased specificity.
Analytical And Bioanalytical Chemistry, 379(7-8):974–981.
PubMed PMID: 15103448.   PDF

2003 Blank K, Mai T, Gilbert I, Schiffmann S, Rankl J, Zivin R, Tackney C, Nicolaus T, Spinnler K, Oesterhelt F, Benoit M, Clausen-Schaumann H and Gaub HE,
A force-based protein biochip.
Proceedings Of The National Academy Of Sciences Of The United States Of America, 100(20):11,356–11360
PubMed PMID: 12975526;   PDF

2002 Benoit M.
Cell adhesion measured by force spectroscopy on living cells.
Methods Cell Biol;68:91-114.
PubMed PMID: 12053742.   PDF

2002 Benoit M and Gaub HE
Measuring cell adhesion forces with the atomic force microscope at the molecular level.
Cells Tissues Organs, 172(3):174–189
PubMed PMID: 12476047.   PDF

2000 Benoit M, Gabriel D, Gerisch G and Gaub HE,
Discrete interactions in cell adhesion measured by single-molecule force spectroscopy.
Nature Cell Biology, 2(6):313–317
PubMed PMID: 10854320.   PDF

2000 Dettmann W, Grandbois M, Andre S, Benoit M, Wehle A, Kaltner H, Gabius H and Gaub HE,
Differences in zero-force and force-driven kinetics of ligand dissociation from beta-galactoside-specific proteins (plant and animal lectins, immunoglobulin G) monitored by plasmon resonance and dynamic single molecule force microscopy.
Archives Of Biochemistry And Biophysics, 383(2):157–170
PubMed PMID: 11185549.   PDF

2000 Grandbois M, Dettmann W, Benoit M and Gaub HE,
Affinity imaging of red blood cells using an atomic force microscope.
Journal Of Histochemistry & Cytochemistry, 48(5):719–724
PubMed PMID: 10769056.   PDF

1998 Thie M, Rospel R, Dettmann W, Benoit M, Ludwig M, Gaub HE and Denker H,
Interactions between trophoblast and uterine epithelium: monitoring of adhesive forces.
Human Reproduction, 13(11):3211–3219
PubMed PMID: 9853883.   PDF

1997 Benoit M, Holstein T and Gaub HE,
Lateral forces in AFM imaging and immobilization of cells and organelles.
European Biophysics Journal With Biophysics Letters, 26(4):283–290.
PubMed PMID: 17838043.   PDF

1994 Holstein TW, Benoit M, Herder vG, Wanner G, David CN, Gaub HE
Fibrous Mini-Collagens in Hydra Nematocysts.
Science, 265(5170):402-4
PMID:17838043   PDF