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Mapping and Modeling the Nanomechanics of Bare and Protein-Coated Lipid Nanotubes

Guillaume Lamour1,*, Antoine Allard1,2, Juan Pelta1, Sid Labdi1, Martin Lenz3, and Clément Campillo1,†

  • 1LAMBE, Université d’Evry, CNRS, CEA, Université Paris-Saclay, 91025, Evry-Courcouronnes, France
  • 2Physico Chimie Curie, CNRS UMR168, Institut Curie, F-75005 Paris, France
  • 3LPTMS, CNRS, Université Paris-Sud, Université Paris-Saclay, 91405 Orsay, France

  • *Corresponding author. guillaume.lamour@univ-evry.fr
  • †Corresponding author. clement.campillo@univ-evry.fr
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Phys. Rev. X 10, 011031 – Published 11 February, 2020

DOI: https://doi.org/10.1103/PhysRevX.10.011031

Abstract

Membrane nanotubes are continuously assembled and disassembled by the cell to generate and dispatch transport vesicles, for instance, in endocytosis. While these processes crucially involve the ill-understood local mechanics of the nanotube, existing micromanipulation assays only give access to its global mechanical properties. Here we develop a new platform to study this local mechanics using atomic force microscopy (AFM). On a single coverslip we quickly generate millions of substrate-bound nanotubes, out of which dozens can be imaged by AFM in a single experiment. A full theoretical description of the AFM tip-membrane interaction allows us to accurately relate AFM measurements of the nanotube heights, widths, and rigidities to the membrane bending rigidity and tension, thus demonstrating our assay as an accurate probe of nanotube mechanics. We reveal a universal relationship between nanotube height and rigidity, which is unaffected by the specific conditions of attachment to the substrate. Moreover, we show that the parabolic shape of force-displacement curves results from thermal fluctuations of the membrane that collides intermittently with the AFM tip. We also show that membrane nanotubes can exhibit high resilience against extreme lateral compression. Finally, we mimic in vivo actin polymerization on nanotubes and use AFM to assess the induced changes in nanotube physical properties. Our assay may help unravel the local mechanics of membrane-protein interactions, including membrane remodeling in nanotube scission and vesicle formation.

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Popular Summary

Lipid bilayers make membranes that delineate the boundaries of living cells or intracellular compartments, and they are continuously remodeled by various types of proteins. In particular, membranes can form cylinders called nanotubes with diameters of 20 to 200 nm. In the cell, nanotubes are cut by specialized proteins to form vesicles, which are then used to transport molecules to a different location of the same cell. The physical mechanisms by which nanotubes assemble and disassemble are unclear because of a lack of experimental techniques to map their morphology and mechanics at the nanoscale. We remedy this problem by developing a new platform to study nanotubes and biological membranes using atomic force microscopy.

We locally probe the mechanics and topography of these soft biological nano-objects by touching their surface with a nanometric indenter: We poke the nanotube surface with the atomic force microscope tip and record force-indentation curves. From these curves, we derive the tube morphology as well as the local tension and bending rigidity of the lipid membrane. Moreover, we test our approach on nanotubes covered by proteins—a network of the cytoskeletal protein actin—and show how actin networks enlarge and stiffen nanotubes.

Our new atomic force microscopy platform may be useful for deciphering the details of nanotube remodeling by proteins and thereby help researchers understand the physics underlying some fundamental cellular processes.

Article Text

References (52)

  1. K. McCoy-Simandle, S. J. Hanna, and D. Cox, Exosomes and Nanotubes: Control of Immune Cell Communication, Int. J. Biochem. Cell Biol. 71, 44 (2016).
  2. A. Roux, The Physics of Membrane Tubes: Soft Templates for Studying Cellular Membranes, Soft Matter 9, 6726 (2013).
  3. P. Bassereau et al., The 2018 Biomembrane Curvature and Remodeling Roadmap, J. Phys. D 51, 343001 (2018).
  4. L. M. Westrate, J. E. Lee, W. A. Prinz, and G. K. Voeltz, Form Follows Function: The Importance of Endoplasmic Reticulum Shape, Annu. Rev. Biochem. 84, 791 (2015).
  5. M. A. De Matteis and A. Luini, Exiting the Golgi Complex, Nat. Rev. Mol. Cell Biol. 9, 273 (2008).
  6. S. Morlot, V. Galli, M. Klein, N. Chiaruttini, J. Manzi, F. Humbert, L. Dinis, M. Lenz, G. Cappello, and A. Roux, Membrane Shape at the Edge of the Dynamin Helix Sets Location and Duration of the Fission Reaction, Cell 151, 619 (2012).
  7. J. Schoneberg et al., ATP-Dependent Force Generation and Membrane Scission by ESCRT-III and Vps4, Science 362, 1423 (2018).
  8. M. Simunovic, P. Bassereau, and G. A. Voth, Organizing Membrane-Curving Proteins: The Emerging Dynamical Picture, Curr. Opin. Struct. Biol. 51, 99 (2018).
  9. M. Kaksonen, C. P. Toret, and D. G. Drubin, Harnessing Actin Dynamics for Clathrin-Mediated Endocytosis, Nat. Rev. Mol. Cell Biol. 7, 404 (2006).
  10. J. Lemiere, F. Valentino, C. Campillo, and C. Sykes, How Cellular Membrane Properties Are Affected by the Actin Cytoskeleton, Biochimie 130, 33 (2016).
  11. S. Miserey-Lenkei, G. Chalancon, S. Bardin, E. Formstecher, B. Goud, and A. Echard, Rab and Actomyosin-Dependent Fission of Transport Vesicles at the Golgi Complex, Nat. Cell Biol. 12, 645 (2010).
  12. W. Roemer et al., Actin Dynamics Drive Membrane Reorganization and Scission in Clathrin-Independent Endocytosis, Cell 140, 540 (2010).
  13. G. Smolyakov, B. Thiebot, C. Campillo, S. Labdi, C. Severac, J. Pelta, and E. Dague, Elasticity, Adhesion, and Tether Extrusion on Breast Cancer Cells Provide a Signature of Their Invasive Potential, ACS Appl. Mater. Interfaces 8, 27426 (2016).
  14. A. Karlsson, R. Karlsson, M. Karlsson, A. S. Cans, A. Stromberg, F. Ryttsen, and O. Orwar, Molecular Engineering: Networks of Nanotubes and Containers, Nature (London) 409, 150 (2001).
  15. M. Karlsson, K. Sott, M. Davidson, A. S. Cans, P. Linderholm, D. Chiu, and O. Orwar, Formation of Geometrically Complex Lipid Nanotube-Vesicle Networks of Higher-Order Topologies, Proc. Natl. Acad. Sci. U.S.A. 99, 11573 (2002).
  16. T. Bhatia, J. Agudo-Canalejo, R. Dimova, and R. Lipowsky, Membrane Nanotubes Increase the Robustness of Giant Vesicles, ACS Nano 12, 4478 (2018).
  17. M. Simunovic et al., Friction Mediates Scission of Tubular Membranes Scaffolded by BAR Proteins, Cell 170, 172 (2017).
  18. P. J. Carman and R. Dominguez, BAR Domain Proteins—A Linkage between Cellular Membranes, Signaling Pathways, and the Actin Cytoskeleton, J. Cell Sci. 10, 1587 (2018).
  19. C. Leduc, O. Campas, J. F. Joanny, J. Prost, and P. Bassereau, Mechanism of Membrane Nanotube Formation by Molecular Motors, BBA-Biomembranes 1798, 1418 (2010).
  20. S. Dar, S. C. Kamerkar, and T. J. Pucadyil, A High-Throughput Platform for Real-Time Analysis of Membrane Fission Reactions Reveals Dynamin Function, Nat. Cell Biol. 17, 1588 (2015).
  21. K. Guevorkian, J. Manzi, L. L. Pontani, F. Brochard-Wyart, and C. Sykes, Mechanics of Biomimetic Liposomes Encapsulating an Actin Shell, Biophys. J. 109, 2471 (2015).
  22. C. Prevost, F. C. Tsai, P. Bassereau, and M. Simunovic, Pulling Membrane Nanotubes from Giant Unilamellar Vesicles, J. Vis. Exp. 130, e56086 (2017).
  23. F. Valentino, P. Sens, J. Lemiere, A. Allard, T. Betz, C. Campillo, and C. Sykes, Fluctuations of a Membrane Nanotube Revealed by High-Resolution Force Measurements, Soft Matter 12, 9429 (2016).
  24. N. Delorme and A. Fery, Direct Method to Study Membrane Rigidity of Small Vesicles Based on Atomic Force Microscope Force Spectroscopy, Phys. Rev. E 74, 030901 (2006).
  25. L. Picas, F. Rico, and S. Scheuring, Direct Measurement of the Mechanical Properties of Lipid Phases in Supported Bilayers, Biophys. J. 102, L01 (2012).
  26. E. Schafer, M. Vache, T. T. Kliesch, and A. Janshoff, Mechanical Response of Adherent Giant Liposomes to Indentation with a Conical AFM-Tip, Soft Matter 11, 4487 (2015).
  27. Y. Zhao, K. Tamhane, X. Zhang, L. An, and J. Fang, Radial Elasticity of Self-Assembled Lipid Tubules, ACS Nano 2, 1466 (2008).
  28. A. Colom, L. Redondo-Morata, N. Chiaruttini, A. Roux, and S. Scheuring, Dynamic Remodeling of the Dynamin Helix during Membrane Constriction, Proc. Natl. Acad. Sci. U.S.A. 114, 5449 (2017).
  29. R. Lipowsky, Coupling of Bending and Stretching Deformations in Vesicle Membranes, Adv. Colloid Interface Sci. 208, 14 (2014).
  30. M. Bovellan et al., Cellular Control of Cortical Actin Nucleation, Curr. Biol. 24, 1628 (2014).
  31. N. Morone, T. Fujiwara, K. Murase, R. S. Kasai, H. Ike, S. Yuasa, J. Usukura, and A. Kusumi, Three-Dimensional Reconstruction of the Membrane Skeleton at the Plasma Membrane Interface by Electron Tomography, J. Cell Biol. 174, 851 (2006).
  32. W. Rawicz, K. C. Olbrich, T. McIntosh, D. Needham, and E. Evans, Effect of Chain Length and Unsaturation on Elasticity of Lipid Bilayers, Biophys. J. 79, 328 (2000).
  33. A. Roux, D. Cuvelier, P. Nassoy, J. Prost, P. Bassereau, and B. Goud, Role of Curvature and Phase Transition in Lipid Sorting and Fission of Membrane Tubules, EMBO J. 24, 1537 (2005).
  34. D. R. Daniels, D. Marenduzzo, and M. S. Turner, Stall, Spiculate, or Run Away: The Fate of Fibers Growing towards Fluctuating Membranes, Phys. Rev. Lett. 97, 098101 (2006).
  35. M. Abramowitz, Handbook of Mathematical Functions, with Formulas, Graphs, and Mathematical Tables (Dover Publications, Inc., New York, 1974).
  36. N. C. Gauthier, T. A. Masters, and M. P. Sheetz, Mechanical Feedback between Membrane Tension and Dynamics, Trends Cell Biol. 22, 527 (2012).
  37. A. D. Lieber, S. Yehudai-Resheff, E. L. Barnhart, J. A. Theriot, and K. Keren, Membrane Tension in Rapidly Moving Cells Is Determined by Cytoskeletal Forces, Curr. Biol. 23, 1409 (2013).
  38. C. Simon, V. Caorsi, C. Campillo, and C. Sykes, Interplay between Membrane Tension and the Actin Cytoskeleton Determines Shape Changes, Phys. Biol. 15, 065004 (2018).
  39. V. Caorsi, J. Lemiere, C. Campillo, M. Bussonnier, J. Manzi, T. Betz, J. Plastino, K. Carvalho, and C. Sykes, Cell-Sized Liposome Doublets Reveal Active Tension Build-Up Driven by Acto-Myosin Dynamics, Soft Matter 12, 6223 (2016).
  40. K. R. Cho, Yu Huang, S. Yu, S. Yin, M. Plomp, S. R. Qiu, R. Lakshminarayanan, J. Moradian-Oldak, M.-S. Sy, and J. J. De Yoreo, A Multistage Pathway for Human Prion Protein Aggregation In Vitro: From Multimeric Seeds to Beta-Oligomers and Nonfibrillar Structures, J. Am. Chem. Soc. 133, 8586 (2011).
  41. Z. Ma, D. N. LeBard, S. M. Loverde, K. A. Sharp, M. L. Klein, D. E. Discher, and T. H. Finkel, TCR Triggering by pMHC Ligands Tethered on Surfaces Via Poly(Ethylene Glycol) Depends on Polymer Length, PLoS One 9, e112292 (2014).
  42. L. Johannes and W. Romer, Shiga Toxins—From Cell Biology to Biomedical Applications, Nat. Rev. Microbiol. 8, 105 (2010).
  43. Y. X. Shen, P. O. Saboe, I. T. Sines, M. Erbakan, and M. Kumar, Biomimetic Membranes: A Review, J. Membr. Sci. 454, 359 (2014).
  44. I. Medalsy, U. Hensen, and D. J. Muller, Imaging and Quantifying Chemical and Physical Properties of Native Proteins at Molecular Resolution by Force-Volume AFM, Angew. Chem. Int. Ed. 50, 12103 (2011).
  45. F. Rico, C. M. Su, and S. Scheuring, Mechanical Mapping of Single Membrane Proteins at Submolecular Resolution, Nano Lett. 11, 3983 (2011).
  46. M. Saleem, S. Morlot, A. Hohendahl, J. Manzi, M. Lenz, and A. Roux, A Balance between Membrane Elasticity and Polymerization Energy Sets the Shape of Spherical Clathrin Coats, Nat. Commun. 6, 6249 (2015).
  47. G. Lamour, S. Souès, and A. Hamraoui, Interplay between Long- and Short-Range Interactions Drives Neuritogenesis on Stiff Surfaces, J. Biomed. Mater. Res. A 99A, 598 (2011).
  48. R. E. Ducker, M. T. Montague, and G. J. Leggett, A Comparative Investigation of Methods for Protein Immobilization on Self-Assembled Monolayers Using Glutaraldehyde, Carbodiimide, and Anhydride Reagents, Biointerphases 3, 59 (2008).
  49. J. E. Sader, J. W. M. Chon, and P. Mulvaney, Calibration of Rectangular Atomic Force Microscope Cantilevers, Rev. Sci. Instrum. 70, 3967 (1999).
  50. K. Carvalho, J. Lemiere, F. Faqir, J. Manzi, L. Blanchoin, J. Plastino, T. Betz, and C. Sykes, Actin Polymerization or Myosin Contraction: Two Ways to Build Up Cortical Tension for Symmetry Breaking, Phil. Trans. R. Soc. B 368, 20130005 (2013).
  51. S. Palmgren, P. J. Ojala, M. A. Wear, J. A. Cooper, and P. Lappalainen, Interactions with PIP2, ADP-Actin Monomers, and Capping Protein Regulate the Activity and Localization of Yeast Twinfilin, J. Cell Biol. 155, 251 (2001).
  52. W. Helfrich, Elastic Properties of Lipid Bilayers: Theory and Possible Experiments, Z. Naturforsch. C 28, 693 (1973).

Outline

Information

Phys. Rev. X 10, 011031– Published 11 February, 2020
  • Received 13 September 2019
  • Revised 22 December 2019
  • Accepted 10 January 2020
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Published by the American Physical Society under the terms of the Creative Commons Attribution 4.0 International license. Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI.

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