The role of the bound nucleotide in the polymerization of actin.
Three mucleotides, ATP, ADP, and an unsplit-table analog of ATP (adenylyl imidodiphosphate (AMPPNP)), were bound to monomeric actin, and their effects on the rate and extent of the actin polymerization were studied. The kinetics of polymerization, assayed by the change in OD232, followed a simple exponential curve. The rates of polymerization were equal for bound ATP and AMPPNP; both of which were three to five times faster than the rate for ADP. The concentration of actin monomers in apparent equilibrium with the polymer, G(180 degrees longitude), was determined. Values of G(180 degrees longitude) in 100 mM KCl were found for different nucleotides to be: G-ATP(180 degrees longitude) = 0.7 mu-M, G-AMPPNP(180 degrees longitude) = 0.8 MU-M, and G-ADP(180 degrees longitude) = 3.4 mu-M. The equilibrium constant of the polymerization is given by K = [G(180 degrees longitude)]-minus 1 when no nucleotide is split. The polymerization of actin-ATP is more complex due to the splitting of the nucleotide and our data require that this polymerization involves more than one step. The kinetic parameters for the polymerization of actin-ATP can be explained by a simple scheme in which the nucleotide dephosphorylation occurs in a step following the polymerization step. The conclusions are: (1) the binding of ATP to actin monomer promotes polymerization slightly more than the binding of ADP, (2) actin bound ATP provides less than 4 kJ/mol of free energy to promote polymerization, and (3) the dephosphorylation of the nucleotide is not coupled to polymerization.
D000199:Actins; D000227:Adenine Nucleotides; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives; D001665:Binding Sites; D007700:Kinetics; D046911:Macromolecular Substances; D008433:Mathematics; D009994:Osmolar Concentration; D011189:Potassium Chloride; D011485:Protein Binding; D013056:Spectrophotometry, Ultraviolet; D013997:Time Factors
D000199:Actins; D000227:Adenine Nucleotides; D046911:Macromolecular Substances; D000244:Adenosine Diphosphate; D011189:Potassium Chloride; D000255:Adenosine Triphosphate