Aberrant cGMP-binding activity in non-chemotactic Dictyostelium discoideum mutants

The kinetics of cGMP-binding to the major cGMP-binding activity in Dictyostelium were investigated in 10 non-chemotactic mutants (KI mutants; KI-1 approximately 10). A wild-type cell contains about 3000 binding sites with a Kd of 1.5 nM. cGMP may dissociate from these binding sites with fast (F-type) or slow (S-type) kinetics, and DNA has been shown to promote the conversion of F- to S-type of cGMP-binding. The 10 mutants were placed in 4 classes, based on equilibrium and non-equilibrium binding properties and the effect of DNA. Class I mutants (KI-1, 3 and 8) have normal cGMP-binding properties. Class II mutants (KI-2, 6 and 7) show increased Kd values but nearly normal Bmax, normal F/S ratio and normal effects of DNA. Class III mutants (KI-4, 5 and 10) have a strongly decreased Kd and increased Bmax, nearly all binding sites are of the S-type and DNA does not affect the binding; apparently these mutants have a cGMP-binding protein locked in the S-form. cGMP-binding in class IV mutant (KI-9) is normal except that the number of binding sites is increased about 3-fold. The finding of seven mutants with altered cGMP-binding in 10 non-chemotactic mutants suggests that the cGMP-binding activity plays an important role in the chemotactic signal transduction pathway.