title: Simultaneous measurement of rotations of myosin, actin and ADP in a contracting skeletal muscle fiber.
abstract: The rotation of myosin heads and actin were measured simultaneously with an indicator of the enzymatic activity of myosin. To minimize complications due to averaging of signals from many molecules, the signal was measured in a small population residing in a femtoliter volume of a muscle fiber. The onset of rotation was synchronized by a sudden release of caged ATP. The orientation of cross-bridges was measured by anisotropy of recombinant fluorescent regulatory light chains exchanged with native regulatory light chains. The orientation of actin was measured by anisotropy of phalloidin added to actin filaments. The enzymatic activity of myosin was measured by dissociation of fluorescent ADP from the active site. The onset of all three events occurred at the same time. This suggests that in contracting muscle, actin does not move independently of myosin and that ATP hydrolysis is strongly coupled to the rotation of cross-bridges.
authors.affiliation: Department of Molecular Biology and Immunology, University of North Texas, 3500 Camp Bowie Building, Fort Worth, TX 76107, USA.
authors.forename: A A
authors.initials: AA
authors.lastname: Shepard
authors.forename: D
authors.initials: D
authors.lastname: Dumka
authors.forename: I
authors.initials: I
authors.lastname: Akopova
authors.forename: J
authors.initials: J
authors.lastname: Talent
authors.forename: J
authors.initials: J
authors.lastname: Borejdo
mesh_terms: D000199:Actins / Q000737:chemistry* / Q000502:physiology; D000244:Adenosine Diphosphate / Q000737:chemistry* / Q000502:physiology; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000502:physiology; D000818:Animals; D009119:Muscle Contraction / Q000502:physiology*; D018485:Muscle Fibers, Skeletal / Q000737:chemistry* / Q000502:physiology; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000502:physiology; D009218:Myosins / Q000737:chemistry* / Q000502:physiology; D011817:Rabbits; D012399:Rotation; D013997:Time Factors
chemical_list: D000199:Actins; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D009218:Myosins