title: Crossbridge and tropomyosin positions observed in native, interacting thick and thin filaments.
abstract: Tropomyosin movements on thin filaments are thought to sterically regulate muscle contraction, but have not been visualized during active filament sliding. In addition, although 3-D visualization of myosin crossbridges has been possible in rigor, it has been difficult for thick filaments actively interacting with thin filaments. In the current study, using three-dimensional reconstruction of electron micrographs of interacting filaments, we have been able to resolve not only tropomyosin, but also the docking sites for weak and strongly bound crossbridges on thin filaments. In relaxing conditions, tropomyosin was observed on the outer domain of actin, and thin filament interactions with thick filaments were rare. In contracting conditions, tropomyosin had moved to the inner domain of actin, and extra density, reflecting weakly bound, cycling myosin heads, was also detected, on the extreme periphery of actin. In rigor conditions, tropomyosin had moved further on to the inner domain of actin, and strongly bound myosin heads were now observed over the junction of the inner and outer domains. We conclude (1) that tropomyosin movements consistent with the steric model of muscle contraction occur in interacting thick and thin filaments, (2) that myosin-induced movement of tropomyosin in activated filaments requires strongly bound crossbridges, and (3) that crossbridges are bound to the periphery of actin, at a site distinct from the strong myosin binding site, at an early stage of the crossbridge cycle.
authors.affiliation: Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA 01655, USA. Roger.Craig@umassmed.edu
authors.forename: R
authors.initials: R
authors.lastname: Craig
authors.forename: W
authors.initials: W
authors.lastname: Lehman
mesh_terms: D000199:Actins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D008854:Microscopy, Electron; D008954:Models, Biological; D008958:Models, Molecular; D009068:Movement; D009119:Muscle Contraction; D009210:Myofibrils / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D009218:Myosins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D013112:Spiders; D014335:Tropomyosin / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*
chemical_list: D000199:Actins; D014335:Tropomyosin; D000255:Adenosine Triphosphate; D009218:Myosins; D002118:Calcium