Binding of α-actinin to F-actin or to tropomyosin F-actin is a function of both α-actinin concentration and gel structure

We have studied by electron microscopy as well as by measurements of low shear viscosity, rigidity and binding, the effect of α-actinin on the gel formed at 37° C with F-actin and with tropomyosin-decorated F-actin. Contrary to previous reports in the literature, α-actinin at nanomolar concentrations is an efficient actin gelling protein, even at 37° C, provided that the concentration of actin (or of tropomyosin-decorated F-actin) is low (1.2-2.4 μm). The binding of α-actinin to F-actin, as a function of actin concentration, is anomalous. The amount of bound α-actinin increases when actin concentration increases from 0 to 1.2 μm but does not change significantly when actin concentration is further increased up to 48 μm. A similar result is obtained with tropomyosin-decorated F-actin. These observations can be explained by an hypothesis that binding is a function of the α-actinin - F-actin association constant as well as of the rigidity of the gel. When the concentration of actin increases, the rigidity of the gel also increases and more work is required to bring two actin filaments to the reaction distance with α-actinin and, consequently, a larger α-actinin concentration is required to attain the same ratio of bound α-actinin to actin monomers in the filaments.