title: The interaction of adenyl-5'-yl imidodiphosphate and PPi with actomyosin.
abstract: We previously studied the equilibrium binding of ADP, adenyl-5'-yl imidodiphosphate (AMP-PNP), and inorganic pyrophosphate (PPi) to actomyosin-subfragment 1 (acto.S-1) and found that AMP-PNP and PPi bind considerably more weakly to acto.S-1 than does ADP. In this study, we investigated the pre-steady-state kinetics of the binding of AMP-PNP and PPi to acto.S-1 and of S-1.AMP-PNP and S-1.PPi to actin to determine if the pre-steady-state kinetic data are consistent with our previous equilibrium data. We find that the kinetic data are consistent with the equilibrium data and agree with a model in which acto.S-1 forms a collision intermediate with the ATP analog, followed by a slower conformational change to a ternary complex that rapidly dissociates into actin and the S-1.ATP analog. Although this scheme fits the AMP-PNP as well as the PPi data, we find that the isomerization of the collision intermediate to the ternary complex is approximately 10 times faster in the presence of PPi than in the presence of AMP-PNP, which is consistent with previous physiological studies (Schoenberg, M., and Eisenberg, E. (1985) Biophys. J. 48, 863-872).
authors.affiliation: Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: J A
authors.initials: JA
authors.lastname: Biosca
authors.forename: E
authors.initials: E
authors.lastname: Eisenberg
mesh_terms: D000199:Actins / Q000378:metabolism; D000205:Actomyosin / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives*; D000266:Adenylyl Imidodiphosphate / Q000378:metabolism*; D000818:Animals; D011756:Diphosphates / Q000378:metabolism*; D007700:Kinetics; D008433:Mathematics; D009132:Muscles / Q000378:metabolism; D011485:Protein Binding; D011817:Rabbits
chemical_list: D000199:Actins; D011756:Diphosphates; D000266:Adenylyl Imidodiphosphate; D000255:Adenosine Triphosphate; D000205:Actomyosin