title: Myosin isoform determines the conformational dynamics and cooperativity of actin filaments in the strongly bound actomyosin complex.
abstract: We used transient phosphorescence anisotropy to detect the microsecond rotational dynamics of erythrosin-iodoacetamide-labeled actin strongly bound to single-headed fragments of muscle myosin subfragment 1 (S1) and non-muscle myosin V (MV). The conformational dynamics of actin filaments in solution are markedly influenced by the isoform of bound myosin. Both myosins increase the final anisotropy of actin at substoichiometric binding densities, indicating long-range, non-nearest neighbor cooperative restriction of filament rotational dynamics amplitude, but the cooperative unit is larger with MV than with muscle S1. Both myosin isoforms also cooperatively affect the actin filament rotational correlation time, but with opposite effects: muscle S1 decreases rates of intrafilament torsional motion, while binding of MV increases the rates of motion. The cooperative effects on the rates of intrafilament motions correlate with the kinetics of myosin binding to actin filaments such that MV binds more rapidly and muscle myosin binds more slowly to partially decorated filaments than to bare filaments. The two isoforms also differ in their effects on the phosphorescence lifetime of the actin-bound erythrosin iodoacetamide: while muscle S1 increases the lifetime, suggesting decreased aqueous exposure of the probe, MV does not induce a significant change. We conclude that the dynamics and structure of actin in the strongly bound actomyosin complex are determined by the isoform of the bound myosin in a manner likely to accommodate the diverse functional roles of actomyosin in muscle and non-muscle cells.
authors: Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.
authors: Ewa
authors: E
authors: Prochniewicz
authors: Harvey F
authors: HF
authors: Chin
authors: Arnon
authors: A
authors: Henn
authors: Diane E
authors: DE
authors: Hannemann
authors: Adrian O
authors: AO
authors: Olivares
authors: David D
authors: DD
authors: Thomas
authors: Enrique M
authors: EM
authors: De La Cruz
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure*; D000205:Actomyosin / Q000378:metabolism* / Q000648:ultrastructure*; D000595:Amino Acid Sequence; D007700:Kinetics; D008969:Molecular Sequence Data; D009038:Motion; D009218:Myosins / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D020033:Protein Isoforms; D000205:Actomyosin; D009218:Myosins