title: Cryoenzymic studies on myosin: transient kinetic evidence for two types of head with different ATP binding properties.
abstract: The two step tight binding of ATP to myosin, heavy meromyosin and myosin subfragment 1 was investigated, under cryoenzymic conditions by the unlabeled ATP chase method: M + ATP in equilibrium K1 M.ATP k2 in equilibrium k-2 M*.ATP where M is myosin. k-2 is close to zero. In multi-turnover experiments, one obtains the constants for the binding process together with the concentration of ATPase sites. Here the kinetics of the formation of M*.ATP are first order. Inversion of the reagent concentrations (i.e., single-turnover experiments) should give identical kinetics but such experiments often give biphasic curves. This biphasicity depends upon the myosin preparation used and it is directly related to the active site titration. The simplest explanation for these results is one involving 2 sites for ATP: one site hydrolyzes ATP by the Bagshaw-Trentham scheme (tight binding preceding hydrolysis) but the second site binds ATP loosely without significant hydrolysis. This heterogeneity in ATP binding may explain certain difficulties, such as questions concerning the non-identity of the myosin heads and the number of steps involved in nucleotide binding. Attempts were made to determine the cause of the head heterogeneity but these were unsuccessful. We cannot exclude the possibility that the heterogeneity is relevant to muscle contraction.
authors.affiliation: INSERM U128, CNRS, Montpellier, France.
authors.forename: C
authors.initials: C
authors.lastname: Tesi
authors.forename: N
authors.initials: N
authors.lastname: Bachouchi
authors.forename: T
authors.initials: T
authors.lastname: Barman
authors.forename: F
authors.initials: F
authors.lastname: Travers
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D007700:Kinetics; D009218:Myosins / Q000378:metabolism*; D011485:Protein Binding; D011817:Rabbits
chemical_list: D000255:Adenosine Triphosphate; D009218:Myosins