Force measurements on natural membrane nanovesicles reveal a composition-independent, high Young's modulus.

Mechanical properties of nano-sized vesicles made up of natural membranes are crucial to the development of stable, biocompatible nanocontainers with enhanced functional, recognition and sensing capabilities. Here we measure and compare the mechanical properties of plasma and inner membrane nanovesicles ∼80 nm in diameter obtained from disrupted yeast Saccharomyces cerevisiae cells. We provide evidence of a highly deformable behaviour for these vesicles, able to support repeated wall-to-wall compressions without irreversible deformations, accompanied by a noticeably high Young's modulus (∼300 MPa) compared to that obtained for reconstituted artificial liposomes of similar size and approaching that of some virus particles. Surprisingly enough, the results are approximately similar for plasma and inner membrane nanovesicles, in spite of their different lipid compositions, especially on what concerns the ergosterol content. These results point towards an important structural role of membrane proteins in the mechanical response of natural membrane vesicles and open the perspective to their potential use as robust nanocontainers for bioapplications.

[1]  Jinwon Park Sulfatide incorporation effect on mechanical properties of vesicles. , 2010, Colloids and surfaces. B, Biointerfaces.

[2]  P. Reis,et al.  Geometry-induced rigidity in nonspherical pressurized elastic shells. , 2012, Physical review letters.

[3]  Yuekan Jiao,et al.  Accurate height and volume measurements on soft samples with the atomic force microscope. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[4]  D. Lasič Novel applications of liposomes. , 1998, Trends in biotechnology.

[5]  Peter Lenz,et al.  Elastic properties and mechanical stability of chiral and filled viral capsids. , 2008, Physical review. E, Statistical, nonlinear, and soft matter physics.

[6]  Howard A Stone,et al.  Mechanics of surface area regulation in cells examined with confined lipid membranes , 2011, Proceedings of the National Academy of Sciences.

[7]  G. Dietler,et al.  Probing nanomechanical properties from biomolecules to living cells , 2008, Pflügers Archiv - European Journal of Physiology.

[8]  Mauricio G Mateu,et al.  Mechanical properties of viruses analyzed by atomic force microscopy: a virological perspective. , 2012, Virus research.

[9]  C. Laroche,et al.  The effect of osmotic pressure on the membrane fluidity of Saccharomyces cerevisiae at different physiological temperatures , 2001, Applied Microbiology and Biotechnology.

[10]  William S Klug,et al.  Nonlinear finite-element analysis of nanoindentation of viral capsids. , 2007, Physical review. E, Statistical, nonlinear, and soft matter physics.

[11]  Arezki Boudaoud,et al.  Indentation of ellipsoidal and cylindrical elastic shells. , 2012, Physical review letters.

[12]  Xuemei Liang,et al.  Mechanical properties and stability measurement of cholesterol-containing liposome on mica by atomic force microscopy. , 2004, Journal of colloid and interface science.

[13]  A. Herrmann,et al.  Bending and puncturing the influenza lipid envelope. , 2011, Biophysical journal.

[14]  Josep Samitier,et al.  A novel detection strategy for odorant molecules based on controlled bioengineering of rat olfactory receptor I7. , 2007, Biosensors & bioelectronics.

[15]  W S Klug,et al.  Nanoindentation studies of full and empty viral capsids and the effects of capsid protein mutations on elasticity and strength. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[16]  C. Veigel,et al.  Effect of Envelope Proteins on the Mechanical Properties of Influenza Virus* , 2012, The Journal of Biological Chemistry.

[17]  M. S. Turner,et al.  Theoretical model for the formation of caveolae and similar membrane invaginations. , 2003, Biophysical journal.

[18]  D. Reguera,et al.  Direct measurement of phage phi29 stiffness provides evidence of internal pressure. , 2012, Small.

[19]  M. S. Turner,et al.  Budded membrane microdomains as tension regulators. , 2006, Physical review. E, Statistical, nonlinear, and soft matter physics.

[20]  M. Persuy,et al.  Diffusion-controlled deposition of natural nanovesicles containing G-protein coupled receptors for biosensing platforms , 2012 .

[21]  H. Schönherr,et al.  Mechanical properties of block copolymer vesicle membranes by atomic force microscopy , 2009 .

[22]  Marie-Annick Persuy,et al.  Functional expression of olfactory receptors in yeast and development of a bioassay for odorant screening , 2005, The FEBS journal.

[23]  J. Vörös,et al.  Liposome and lipid bilayer arrays towards biosensing applications. , 2010, Small.

[24]  H. Hansma,et al.  Changes in the elastic properties of cholinergic synaptic vesicles as measured by atomic force microscopy. , 1997, Biophysical journal.

[25]  J. Bouwstra,et al.  Vesicles as a tool for transdermal and dermal delivery. , 2005, Drug discovery today. Technologies.

[26]  C. Martini,et al.  Multiscale morphology of organic semiconductor thin films controls the adhesion and viability of human neural cells. , 2010, Biophysical journal.

[27]  Marie-Annick Persuy,et al.  Quantitative assessment of olfactory receptors activity in immobilized nanosomes: a novel concept for bioelectronic nose. , 2006, Lab on a chip.

[28]  C. Théry,et al.  Membrane vesicles as conveyors of immune responses , 2009, Nature Reviews Immunology.

[29]  Horst Vogel,et al.  Investigating cellular signaling reactions in single attoliter vesicles. , 2005, Journal of the American Chemical Society.

[30]  E. Work,et al.  Laboratory techniques in biochemistry and molecular biology , 1969 .

[31]  E. Zinser,et al.  Sterol composition of yeast organelle membranes and subcellular distribution of enzymes involved in sterol metabolism , 1993, Journal of bacteriology.

[32]  W F Heinz,et al.  Spatially resolved force spectroscopy of biological surfaces using the atomic force microscope. , 1999, Trends in biotechnology.

[33]  Samuel A Wickline,et al.  A systematic approach to exosome-based translational nanomedicine. , 2012, Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology.

[34]  H. Kleinman,et al.  Extracellular membrane vesicles from tumor cells promote angiogenesis via sphingomyelin. , 2002, Cancer research.

[35]  Tai Hyun Park,et al.  Nanovesicle-based bioelectronic nose platform mimicking human olfactory signal transduction. , 2012, Biosensors & bioelectronics.

[36]  G. Wuite,et al.  Bacteriophage capsids: tough nanoshells with complex elastic properties. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[37]  F. Sanz,et al.  Nanomechanics of lipid bilayers: heads or tails? , 2010, Journal of the American Chemical Society.

[38]  S. Kohlwein,et al.  Phospholipid synthesis and lipid composition of subcellular membranes in the unicellular eukaryote Saccharomyces cerevisiae , 1991, Journal of bacteriology.

[39]  P. Leclère,et al.  Quantitative Measurement of the Mechanical Contribution to Tapping-Mode Atomic Force Microscopy Images of Soft Materials , 2000 .

[40]  K. Murakoshi,et al.  Tuning the dynamics and molecular distribution of the self-spreading lipid bilayer. , 2008, Physical chemistry chemical physics : PCCP.

[41]  Fausto Sanz,et al.  Nanomechanics of lipid bilayers by force spectroscopy with AFM: a perspective. , 2010, Biochimica et biophysica acta.

[42]  Ger J.A. Arkesteijn,et al.  Quantitative and qualitative flow cytometric analysis of nanosized cell-derived membrane vesicles , 2011, Nanomedicine: Nanotechnology, Biology and Medicine.

[43]  G. Vancso,et al.  pH dependent elasticity of polystyrene-block-poly(acrylic acid) vesicle shell membranes by atomic force microscopy. , 2011, Macromolecular rapid communications.

[44]  Y. Obata,et al.  Novel ultra-deformable vesicles entrapped with bleomycin and enhanced to penetrate rat skin. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[45]  Xuemei Liang,et al.  Probing small unilamellar EggPC vesicles on mica surface by atomic force microscopy. , 2004, Colloids and surfaces. B, Biointerfaces.

[46]  Sandor Kasas,et al.  Deformation and height anomaly of soft surfaces studied with an AFM , 1993 .

[47]  M. Morilla,et al.  Topical and mucosal liposomes for vaccine delivery. , 2011, Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology.

[48]  J. Gómez‐Herrero,et al.  WSXM: a software for scanning probe microscopy and a tool for nanotechnology. , 2007, The Review of scientific instruments.

[49]  Udo Seifert,et al.  Configurations of fluid membranes and vesicles , 1997 .

[50]  C. Laroche,et al.  Phase transitions as a function of osmotic pressure in Saccharomyces cerevisiae whole cells, membrane extracts and phospholipid mixtures. , 2005, Biochimica et biophysica acta.

[51]  Alexander M Seifalian,et al.  The application of exosomes as a nanoscale cancer vaccine , 2010, International journal of nanomedicine.

[52]  I. Fichtner,et al.  Treatment of Experimental Brain Metastasis with MTO-Liposomes: Impact of Fluidity and LRP-Targeting on the Therapeutic Result , 2012, Pharmaceutical Research.

[53]  R. Haguenauer‐Tsapis,et al.  Deubiquitination Step in the Endocytic Pathway of Yeast Plasma Membrane Proteins: Crucial Role of Doa4p Ubiquitin Isopeptidase , 2001, Molecular and Cellular Biology.

[54]  L. Bednarz,et al.  Direct measurement and control of peak tapping forces in atomic force microscopy for improved height measurements , 2011 .

[55]  H. S. Marinho,et al.  Gel Domains in the Plasma Membrane of Saccharomyces cerevisiae , 2010, The Journal of Biological Chemistry.

[56]  S. Lim,et al.  Exosomes for drug delivery - a novel application for the mesenchymal stem cell. , 2013, Biotechnology advances.

[57]  A. Fery,et al.  Direct method to study membrane rigidity of small vesicles based on atomic force microscope force spectroscopy. , 2006, Physical review. E, Statistical, nonlinear, and soft matter physics.

[58]  A. Nakano,et al.  The plasma membrane of Saccharomyces cerevisiae: structure, function, and biogenesis. , 1995, Microbiological reviews.