Abstract
Mechanical and structural properties of K8/K18 and vimentin intermediate filament (IF) networks have been investigated using bulk mechanical rheometry and optical microrheology including diffusing wave spectroscopy and multiple particle tracking. A high elastic modulus G 0 at low protein concentration c, a weak concentration dependency of G 0 (G 0∼c 0.5±0.1) and pronounced strain stiffening are found for these systems even without external crossbridgers. Strong attractive interactions among filaments are required to maintain these characteristic mechanical features, which have also been reported for various other IF networks. Filament assembly, the persistence length of the filaments and the network mesh size remain essentially unaffected when a nonionic surfactant is added, but strain stiffening is completely suppressed, G 0 drops by orders of magnitude and exhibits a scaling G 0∼c 1.9±0.2 in agreement with microrheological measurements and as expected for entangled networks of semi-flexible polymers. Tailless K8Δ/K18ΔT and various other tailless filament networks do not exhibit strain stiffening, but still show high G 0 values. Therefore, two binding sites are proposed to exist in IF networks. A weaker one mediated by hydrophobic amino acid clusters in the central rod prevents stretched filaments between adjacent cross-links from thermal equilibration and thus provides the high G 0 values. Another strong one facilitating strain stiffening is located in the tail domain with its high fraction of hydrophobic amino acid sequences. Strain stiffening is less pronounced for vimentin than for K8/K18 due to electrostatic repulsion forces partly compensating the strong attraction at filament contact points.
Highlights
Mechanical properties of metazoan cells are determined by three distinct types of filament systems: F-actin, intermediate filaments (IFs) and microtubules [1]
Common features of pure IF networks at physiological conditions are the pronounced elasticity at small deformations as characterized by a frequency independent storage modulus G0 and the weak dependence of G0 on protein concentration found for vimentin [5,6,7], desmin [7] and keratin [6]
Mesh size and homogeneity of K8/K18 filament networks are directly obtained from multiple particle tracking (MPT) experiments
Summary
Mechanical properties of metazoan cells are determined by three distinct types of filament systems: F-actin, intermediate filaments (IFs) and microtubules [1]. Common features of pure IF networks at physiological conditions are the pronounced elasticity at small deformations as characterized by a frequency independent storage modulus G0 and the weak dependence of G0 on protein concentration found for vimentin [5,6,7], desmin [7] and keratin [6]. This property strongly disagrees with the results obtained for actin networks [5,8] and contradict existing theoretical models for networks of flexible or semi-flexible polymers [8,9,10,11]. At protein concentrations above 1.5 g/l or at divalent ions concentrations above 2 mM, G0 and its scaling with protein concentration are similar to what is expected from the above mentioned theoretical models [12,13]
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