Active elastic network: Cytoskeleton of the red blood cell Nir S. Gov Department of Chemical Physics, The Weizmann Institute of Science, P.O.B. 26, Rehovot, Israel 76100 Received 25 September 2006; published 19 January 2007 In red blood cells there is a cortical cytoskeleton; a two-dimensional elastic network of membrane-attached proteins. We describe, using a simple model, how the metabolic activity of the cell, through the consumption ofATP, controls the stiffness of this elastic network. The unusual mechanical property of active strain softening is described and compared to experimental data. As a by-product of this activity there is also an active contribution to the amplitude of membrane fluctuations. We model this membrane as a field of independent “curvature motors,” and calculate the spectrum of active fluctuations. We find that the active cytoskeleton contributes to the amplitude of the membrane height fluctuations at intermediate wavelengths, as observed experimentally. DOI: 10.1103/PhysRevE.75.011921 PACS numbers: 87.68.+z, 83.60.-a, 87.16.Ac, 87.17.-d I. INTRODUCTION The red-blood cell RBChas a cortical cytoskeleton, which is a two-dimensional network of cross-linked elastic proteins that are attached to and cover the entire inner sur- face of the cell membrane 1,2. Such a cortical cytoskeleton gives the RBC the elasticity to survive repeated deformations 3. The outstanding problem of understanding the physical properties of the RBC cytoskeleton has been to reconcile the measurements of the elasticity of the cell 3, the observed shapes of the RBC 4, and the measurements of the dynamic shape fluctuations of the membrane 5,6. Our model at- tempts to reconcile these observation by showing that they follow naturally from our recently proposed mechanism of active remodeling of the RBC cytoskeleton 7. In this paper we extend our recent theoretical modeling 7,8and calculate several additional features of the active cytoskeleton. iWe calculate the unique effect of strain-softening in this active network. The active dissociations of the network can account for the observed strain-softening of the RBC due to the dis- sociated filaments spending a longer time disconnected when the network is stretched. iiWe then calculate the spectrum of active fluctuations, where we find that these contribute to the membrane height fluctuations at intermediate wave- lengths, as observed experimentally. The RBC cytoskeleton is a self-assembled network of flexible spectrin filaments that are end-attached to node com- plexes Fig. 1. On average approximately six spectrin fila- ments attach to each actin node, mainly through the protein- 4.1 1,9, with a specific chemical affinity of order 7k B T. The flexible spectrin filaments are much longer 200 nm than the distance between the nodes that they connect R 80-100 nm, Fig. 1a, and can be treated as wormlike chains with an average end-to-end distance of 10: R 0 50- 60 nm. Each filament behaves for small deforma- tionsas a Hookean entropic spring, with a spring constant of 11: k 20-40k B T / R 2 1 10 -5 J/m 2 . At larger extensions a semiflexible chain behaves nonlinearly, and exhibits strain stiffening 12,13and unfolding 14. Note that the spectrin filaments are additionally connected to the membrane through the ankyrin-band-3 complex at random locations along the filaments, in between the actin-band-4.1 complexes at their ends 1. It is the connections at the actin nodes that determine the overall connectivity of the network, and there- fore dominate in controlling its elastic properties. We will therefore, also for simplicity of modeling, deal here with the active processes occurring at the filaments ends alone. The elasticity of such a two-dimensional network, at- tached to the fluid bilayer, can be modeled as a static elastic network, with fixed connectivity 4,11,15. Nevertheless, ob- servations of membrane fluctuations 6led to the specula- tion that ATP-induced phosphorylation of cytoskeleton pro- teins induces an overall softening. More recently, numerical simulations 4recover the observed discocyte shape only FIG. 1. A schematic illustration of our model of ATP-induced spectrin dissociation. The main parts of the figure show a side view of the RBC membrane, while the small parts show a schematic top view of the triangular cytoskeleton network. aThe fully connected network, showing the balance between the stretched spectrin and curved bilayer. bThe dissociated spectrin releases its stored ten- sion. The phosphorylated node is indicated by the empty circle. PHYSICAL REVIEW E 75, 011921 2007 1539-3755/2007/751/0119216©2007 The American Physical Society 011921-1