title: The ultrastructure of fibronectin fibers pulled from a protein monolayer at the air-liquid interface and the mechanism of the sheet-to-fiber transition.
abstract: Fibronectin is a globular protein that circulates in the blood and undergoes fibrillogenesis if stretched or under other partially denaturing conditions, even in the absence of cells. Stretch assays made by pulling fibers from droplets of solutions containing high concentrations of fibronectin have previously been introduced in mechanobiology, particularly to ask how bacteria and cells exploit the stretching of fibronectin fibers within extracellular matrix to mechano-regulate its chemical display. Our electron microscopy analysis of their ultrastructure now reveals that the manually pulled fibronectin fibers are composed of densely packed lamellar spirals, whose interlamellar distances are dictated by ion-tunable electrostatic interactions. Our findings suggest that fibrillogenesis proceeds via an irreversible sheet-to-fiber transition as the fibronectin sheet formed at the air-liquid interface of the droplet is pulled off by a sharp tip. This far from equilibrium process is driven by the externally applied force, interfacial surface tension, shear-induced fibronectin self-association, and capillary force-induced buffer drainage. The ultrastructural characterization is then contrasted with previous FRET studies that characterized the molecular strain within these manually pulled fibers. Particularly relevant for stretch-dependent binding studies is the finding that the interior fiber surfaces are accessible to nanoparticles smaller than 10 nm. In summary, our study discovers the underpinning mechanism by which highly hierarchically structured fibers can be generated with unique mechanical and mechano-chemical properties, a concept that might be extended to other bio- or biomimetic polymers.
authors.affiliation: Laboratory of Applied Mechanobiology, Vladimir-Prelog-Weg 4, ETH Zurich, Switzerland.
authors.forename: Maria
authors.initials: M
authors.lastname: Mitsi
authors.affiliation: SCOPEM - Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland.
authors.forename: Stephan
authors.initials: S
authors.lastname: Handschin
authors.affiliation: Laboratory of Applied Mechanobiology, Vladimir-Prelog-Weg 4, ETH Zurich, Switzerland.
authors.forename: Isabel
authors.initials: I
authors.lastname: Gerber
authors.affiliation: Laboratory of Applied Mechanobiology, Vladimir-Prelog-Weg 4, ETH Zurich, Switzerland.
authors.forename: Ruth
authors.initials: R
authors.lastname: Schwartländer
authors.affiliation: Laboratory of Applied Mechanobiology, Vladimir-Prelog-Weg 4, ETH Zurich, Switzerland.
authors.forename: Enrico
authors.initials: E
authors.lastname: Klotzsch
authors.affiliation: SCOPEM - Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland.
authors.forename: Roger
authors.initials: R
authors.lastname: Wepf
authors.affiliation: Laboratory of Applied Mechanobiology, Vladimir-Prelog-Weg 4, ETH Zurich, Switzerland. Electronic address: viola.vogel@hest.ethz.ch.
authors.forename: Viola
authors.initials: V
authors.lastname: Vogel
mesh_terms: D000388:Air / Q000032:analysis; D001696:Biomechanical Phenomena; D005353:Fibronectins / Q000737:chemistry / Q000302:isolation & purification / Q000648:ultrastructure*; D006801:Humans; D008854:Microscopy, Electron; D009994:Osmolar Concentration; D010539:Permeability; D012996:Solutions / Q000737:chemistry; D013499:Surface Properties
chemical_list: D005353:Fibronectins; D012996:Solutions
keywords: Electron microscopy; Fibrillogenesis; Fibronectin; Kinetically trapped supramolecular system; Monolayer-to-lamella transition; Sheet-to-fiber transition