title: Close proximity of myosin loop 3 to troponin determined by triangulation of resonance energy transfer distance measurements.
abstract: Cooperative activation of the thin filament is known to be influenced by the tight binding of myosin to actin, but the molecular mechanism underlying this contribution of myosin is not well understood. To better understand the structural relationship of myosin with the regulatory troponin complex, resonance energy transfer measurements were used to map the location of troponin relative to a neighboring myosin bound to actin using atomic models. Using a chicken troponin T isoform that contains a single cysteine near the binding interface between troponins T, I, and C, this uniquely labeled cysteine on troponin was found to be remarkably near loop 3 of myosin. This loop has previously been localized near the actin and myosin interface by chemical cross-linking methods, but its functional contributions have not been established. The implications of this close proximity are examined by molecular modeling, which suggests that only restricted conformations of actomyosin can accommodate the presence of troponin at this location near the cross-bridge. This potential for interaction between troponin and myosin heads that bind near it along the thin filament raises the possibility of models in which direct myosin and troponin interactions may play a role in the regulatory mechanism.
authors.affiliation: University of North Texas, P.O. Box 305220, Denton, Texas 76203-5220, USA.
authors.forename: Dipesh A
authors.initials: DA
authors.lastname: Patel
authors.forename: Douglas D
authors.initials: DD
authors.lastname: Root
mesh_terms: D008841:Actin Cytoskeleton / Q000737:chemistry / Q000378:metabolism; D000199:Actins / Q000032:analysis / Q000378:metabolism; D000205:Actomyosin / Q000032:analysis / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000032:analysis / Q000378:metabolism; D000818:Animals; D002645:Chickens; D002918:Chymotrypsin / Q000494:pharmacology; D003545:Cysteine / Q000737:chemistry; D019281:Dimerization; D031541:Fluorescence Resonance Energy Transfer; D008954:Models, Biological; D008958:Models, Molecular; D018482:Muscle, Skeletal / Q000737:chemistry; D009218:Myosins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D010206:Papain / Q000494:pharmacology; D020033:Protein Isoforms / Q000032:analysis / Q000378:metabolism; D017433:Protein Structure, Secondary; D011817:Rabbits; D014336:Troponin / Q000032:analysis* / Q000378:metabolism
chemical_list: D000199:Actins; D020033:Protein Isoforms; D014336:Troponin; D000255:Adenosine Triphosphate; D000205:Actomyosin; D002918:Chymotrypsin; C117608:alpha-chymotrypsin; D010206:Papain; D009218:Myosins; D003545:Cysteine