title: Kinetic mechanism of myofibril ATPase.
abstract: The kinetic mechanism of myofibril ATPase was investigated using psoas and mixed back muscle over a range of ionic strengths. Myofibrils were labeled with pyrene iodoacetamide to measure the rate constants for the binding of ATP and formation of the weakly attached state. The velocity of shortening was measured by stopping the contraction at various times by mixing with pH 4.5 buffer. The transient and steady-state rates of ATP hydrolysis were measured by the quench flow method. The results fitted the kinetic scheme [formula: see text] The rate constants (or equilibrium constants for steps 1 and 6) were obtained for the six steps. k5 was calculated from the KM for shortening velocity, K1, and k2. The rate constants were essentially equal for myofibrils and acto-S-1 at low ionic strength. Increasing the ionic strength up to 100 mM in NaCl increased the rate of the hydrolysis step and the size of the phosphate burst and the effective rate of product release became the rate-limiting step. The step calculated from the velocity of shortening, k5, and k2 is 15 nm, based on a model in which step 4 is the force-generating step.
authors.affiliation: Department of Molecular Genetics and Cell Biology, University of Chicago, Illinois 60637.
authors.forename: Y Z
authors.initials: YZ
authors.lastname: Ma
authors.forename: E W
authors.initials: EW
authors.lastname: Taylor
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D006868:Hydrolysis; D066298:In Vitro Techniques; D007700:Kinetics; D009119:Muscle Contraction / Q000502:physiology; D009210:Myofibrils / Q000201:enzymology*; D009218:Myosins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism / Q000494:pharmacology; D011721:Pyrenes
chemical_list: D010710:Phosphates; D011721:Pyrenes; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; C030984:pyrene; D009218:Myosins