Probe diffusion in cross-linked actin gels.
The diffusion of monodisperse polystyrene latex spheres (PLS) in column-purified 0.65 mg/mL actin solutions, polymerized with 100 mM KCl in the absence and presence of a cross-linker, actin binding protein (ABP), has been studied using dynamic light scattering. Measurements over a wide range of scattering angles from 90 degrees to 8 degrees, corresponding to inverse scattering vector probing distances of about 40-400 nm, respectively, give a measure of both the fraction of PLS mobile over the probing distance (from the normalized time autocorrelation function amplitude) and the average diffusion coefficient of the mobile PLS. Both 100- and 500-nm diameter PLS are fully mobile in polymerized actin solutions over distances of less than 100 nm, as reported previously (Newman, J., Schick, K. S. & Gukelberger, G. Biophys. J. 53, 573a, and Newman, J., Mroczka, N. & Schick, K. L. Biopolymers, 28, 655-666). At increasing probing distances, or when ABP is added at molar ratios of 1:750 or 1:150, greater fractions of the PLS are immobilized, up to almost 99% at the conditions of a 400-nm probing distance with 500-nm probes and at a ratio of 1:150 added ABP to actin. The degree of immobilization correlated well with the amount of added ABP, the size of the PLS, and the probing distance. At increasing probe distances, as the degree of immobilization increases, the remaining mobile fraction of PLS has an increasing average diffusion coefficient. These results suggest a range of pore sizes in the actin gels with a mean size of a few hundred nanometers.
Newman
J
J
Schick
K L
KL
Zaner
K S
KS
D000199:Actins; D000818:Animals; D003432:Cross-Linking Reagents; D004058:Diffusion; D005782:Gels; D008027:Light; D008840:Microfilament Proteins; D011817:Rabbits; D012542:Scattering, Radiation
D000199:Actins; D003432:Cross-Linking Reagents; D005782:Gels; D008840:Microfilament Proteins