title: Interaction of two heads of myosin with F-actin: binding of H-meromyosin with F-actin in the absence of nucleotide.
abstract: The bindings of S-1 and the two heads of HMM with pyrene-labeled F-actin were studied using the change in light-scattering intensity or that in the fluorescence intensity of the pyrenyl group. At low ionic strength (50 mM KCl), both S-1 and HMM became bound tightly with F-actin (Kd less than 0.1 microM) and both heads of HMM became bound to F-actin. The affinities of S-1 and HMM for F-actin decreased with increasing KCl concentration. In 1 M KCl, the Kd values of S-1 and HMM for F-actin were 11 and 0.58 microM, respectively. Thus, HMM was bound to F-actin 19 times more tightly than S-1. We compared the extent of binding of HMM to F-actin measured by a centrifugation method with that measured by the fluorescence change of pyrenyl-group, and found that even in 1 M KCl, HMM became bound to F-actin with a two-headed attachment. We measured the kinetics of binding and dissociation of acto-S-1 and acto-HMM from the time course of the change in light-scattering intensity after mixing S-1 or HMM with F-actin at 1 M KCl and that after mixing 1 M KCl with acto-S-1 or acto-HMM formed at low ionic strength.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Biology, Faculty of Science, Osaka University.
authors.forename: M
authors.initials: M
authors.lastname: Miyata
authors.forename: T
authors.initials: T
authors.lastname: Arata
authors.forename: A
authors.initials: A
authors.lastname: Inoue
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D007700:Kinetics; D009218:Myosins / Q000378:metabolism*; D011485:Protein Binding; D011721:Pyrenes; D011817:Rabbits
chemical_list: D000199:Actins; D011721:Pyrenes; C030984:pyrene; D009218:Myosins