Magnetic tweezers are applied to study the enforced motion of single actin filaments in entangled actin networks to gain insight into friction-mediated entanglement in semiflexible macromolecular networks. Magnetic beads are coupled to one chain end of test filaments, which are pulled by 5 to 20 pN force pulses through entangled solutions of nonlabeled actin, the test filaments thus acting as linear force probes of the network. The transient filament motion is analyzed by microfluorescence, and the deflectionversus-time curves of the beads are evaluated in terms of a mechanical equivalent circuit to determine viscoelastic parameters, which are then interpreted in terms of viscoelastic moduli of the network. We demonstrate that the frictional coefficient characterizing the hydrodynamic coupling of the filaments to the surrounding network is much higher than predicted by the tube model, suggesting that friction-mediated interfilament coupling plays an important role in the entanglement of non-cross-linked actin networks. Furthermore, the local tube width along the filament contour (measured in terms of the root-mean-square displacement characterizing the lateral Brownian motion of the test filament) reveals strong fluctuations that can lead to transient local pinching of filaments. N etworks of filamentous actin (F-actin) are of great interest from the point of view of both cell biology and polymer physics and have thus been the subject of intense experimental and theoretical studies. On the one hand, actin is a major structural component of the intracellular scaffold (the cytoskeleton) and plays a key role in various cellular processes, such as cell locomotion (1, 2), the transport processes within cells (3), or the control of cell adhesion on surfaces (4). To fulfill this multiplicity of tasks, nature uses a large number of helper proteins. These include severing proteins by which the length of actin filaments can be manipulated, monomer-sequestering proteins that allow the control of the polymer concentration, and finally a manifold of cross-linker proteins enabling the generation of randomly organized gels or arrangements of bundles acting as cell-stabilizing fibers or cables for intracellular transport (5).On the other hand, F-actin is a prototype of a semiflexible polymer. Highly versatile models of entangled and cross-linked networks of semiflexible macromolecules can be designed by controlling the structure through the manifold of manipulating proteins to study the distinct physical properties of this particular class of polymer networks. Such semiflexible polymer networks exhibit outstanding viscoelastic features, which are determined by a subtle interplay of entropic and enthalpic contributions to the elastic free energy of the individual filament (6-12).One distinct advantage of actin as model polymer is the large contour length (typically in the 10-m range) (13) and persistence length (L p Ӎ17 m) (10, 14, 15) enabling the design of networks with mesh sizes in the optical wavelength regime. Therefore, t...
This paper deals with correlations between the viscoelastic impedance of entangled actin networks and the slow conformational dynamics and diffusive motions of single filaments. The single filament dynamics is visualized and analysed by analysing the Brownian motion of attached colloidal beads, which enables independent measurements of characteristic viscoelastic response times such as the entanglement and reptation times. We further studied the frequency-dependent viscoelastic impedance of active actin-heavy-meromyosin II networks by magnetic-tweezers microrheometry to gain insight into the effect of such highly dynamic and force-generating crosslinkers (exhibiting bond lifetimes of less than 1 s) on the rheological properties. We show that at high frequencies (higher than 1 Hz) the viscoelastic loss modulus is slightly increased relative to the entangled network (associated with an increase in the energy dissipated during mechanical excitations), while at low frequencies the plateau of the impedance spectrum becomes more pronounced as a consequence of the cross-linking of the network and the suppression of the terminal regime. Our data provide evidence that the myosin motor protein may play a role as softener of the actin cortex, enabling the adaptive reduction of the yield stress of cells and thus facilitating cellular deformations.
This paper deals with correlations between the viscoelastic impedance of entangled actin networks and the slow conformational dynamics and diffusive motions of single filaments. The single filament dynamics is visualized and analysed by analysing the Brownian motion of attached colloidal beads, which enables independent measurements of characteristic viscoelastic response times such as the entanglement and reptation times. We further studied the frequency-dependent viscoelastic impedance of active actin-heavy-meromyosin II networks by magnetic-tweezers microrheometry to gain insight into the effect of such highly dynamic and force-generating crosslinkers (exhibiting bond lifetimes of less than 1 s) on the rheological properties. We show that at high frequencies (higher than 1 Hz) the viscoelastic loss modulus is slightly increased relative to the entangled network (associated with an increase in the energy dissipated during mechanical excitations), while at low frequencies the plateau of the impedance spectrum becomes more pronounced as a consequence of the cross-linking of the network and the suppression of the terminal regime. Our data provide evidence that the myosin motor protein may play a role as softener of the actin cortex, enabling the adaptive reduction of the yield stress of cells and thus facilitating cellular deformations.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
customersupport@researchsolutions.com
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.