The Kinetics of the Exchange of G‐Actin‐Bound 1: N6‐Ethenoadenosine 5′‐Triphosphate with ATP as Followed by Fluorescence

1: N6-Ethenoadenosine 5'-triphosphate (epsilonATP), a fluorescent analog of ATP, binds to monomeric actin with a binding constant which is only about 5 times smaller than that of ATP. The spectroscopic changes which occur when epsilonATP binds to actin are studied and used to monitor the kinetics of nucleotide exchange. The first-order rate constant which is measured at a large excess of ATP over epsilonATP strongly depends on the ATP and Ca+ concentrations. This finding is explained by a mechanism in which the nucleotide dissociates much more easily from Ca2+-free than from Ca2+-bound actin. Of special interest is the temperature dependence of the dissociation rate constant. The Arrhenius plot shows a sharp bend near 24 degrees C.