Selectivity of interaction of univalent cations with mammalian ribosomes studied by equilibrium dialysis in the presence of the K+ analogue, 204Tl+.

The interaction of K+ with mammalian ribosomes was studied by equilibrium dialysis and compared with that of other univalent cations. The heavy K+ analogue, Tl+, binds more firmly than K+ to ribosomes and, unlike K+, has a practically useful isotope. With 204Tl+ as a marker of K+-selective binding the ribosome-cation interaction could be followed down to levels below 0.1 average Tl+-occupied site per ribosome. The Tl+/ribosome ratio varied with the free Tl+ concentration in a multiple way. At high Tl+ saturation Tl+ was easily displaced by Mg2+. With decreasing Tl+ saturation the competitive activity of Mg++ was strikingly reduced, indicating that Tl+ and Mg++ compete with different efficiency for different classes of sites. The experiments on univalent cations were performed at 1.5 mM Mg2+ under two complementary conditions: (1) Ribosomes were pretreated with 5 x 10(-2), 5 x 10(-3), and 5 x 10(-4) M LiNO3, NaNO3, KNO3, and CsNO3, and then equilibrated with different concentrations of 204TlNO3 in the same buffers. (2) Ribosomes were pretreated with 10(-2), 10(-4), and 10(-6) M 204 TlNO3, and then equilibrated with different concentrations of LiNO3, NaNO3, KNO3, and CsNO3 (displacement experiments). At high Tl+ saturation Na+ and Li+ were about as active as K+ and Cs+ in competing with 204Tl+. With decreasing Tl+ saturation a differentiation occurred in favor of K+ and Cs+, with some preference for K+. It is concluded that ribosomes contain a limited number of sites with pronounced ion specificity. Of physiological cations K+ is most firmly bound to these sites.