Evidence for the non-filamentous aggregation of actin induced by lanthanide ions.

We varied the molar ratio of added lanthanide ion to skeletal muscle actin (M3+/A) and observed their effects on the change in reduced viscosity (Nred) in the presence of polymerizing quantities of salt (0.1 M KC1). Once the concentration of the lanthanide ion exceeds the concentration of the nucleotide present (0.2 mM ATP), we noted that with M3+/A ratios up to 4: (a) there was a sharp peak in the observed Nred above the level achieved by control F-actin; (b) the magnitude of (a) was shown to be a function of the initial G-actin concentration. With an M3+/A ratio of greater than 4 we observed: (i) a sharp fall in the observed Nred; (ii) the formation of an insoluble aggregate of actin; (iii) the formation of (ii) was completely reversed by removal of the M3+; (iv) a complete inhibition of the ATP hydrolysis which always accompanies the G- to F-actin transition; (v) the number of mol of M3+ required to completely inhibit the rise in Nred (above the viscosity of G-actin) was a function of the ionic radii of the 11 lanthanide ions tested; and (vi) the effects described in (i) were not mimicked when the initial protein was in the F form. In the absence of added KCI, divalent cations (e.g. Mg2+) polymerize G-actin but this effect is not mimicked by the addition of the lanthanide ions. However, under these conditions the lanthanide ions cause the formation of an insoluble aggregate of actin. We conclude that with greater than 4 mol of lanthanide ions, G-actin aggregates in a form which contains little or no F-actin and that the lanthanide ion-induced aggregates are therefore different from the Mg2+-induced F-actin paracrystals.