A new procedure for synthesis of 14C-labeled tentoxin [14C-MePhe[(Z)delta]3-tentoxin], with a high specific activity, is described. Binding experiments with CF1 or CF1-epsilon isolated from spinach chloroplast have been carried out using equilibrium dialysis technique. The results show the presence of two classes of binding sites. The association constants of the two major binding sites were derived from non-linear fitting of the binding curves. At 4 degrees C, the first binding site has a value of Ka1 = 8.2 x 10(5) M(-1) in CF1 and 8.7 x 10(5) M(-1) in CF1-epsilon, while the second binding site has lower affinity with Ka2 = 1.5 x 10(4) M(-1) in CF1 and 2.3 x 10(3) M(-1) in CF1-epsilon.