title: The structural dynamics of actin during active interaction with myosin depends on the isoform of the essential light chain.
abstract: We have used time-resolved phosphorescence anisotropy to investigate the effects of essential light chain (ELC) isoforms (A1 and A2) on the interaction of skeletal muscle myosin with actin, to relate structural dynamics to previously reported functional effects. Actin was labeled with a phosphorescent probe at C374, and the myosin head (S1) was separated into isoenzymes S1A1 and S1A2 by ion-exchange chromatography. As previously reported, S1A1 exhibited substantially lower ATPase activity at saturating actin concentrations but substantially higher apparent actin affinity, resulting in a higher catalytic efficiency. In the absence of ATP, each isoenzyme increased actin's final anisotropy cooperatively and to a similar extent, indicating a similar restriction of the amplitude of intrafilament rotational motions in the strong-binding (S) state of actomyosin. In contrast, in the presence of a saturating level of ATP, S1A1 increased actin anisotropy much more than S1A2 and with greater cooperativity, indicating that S1A1 was more effective in restricting actin dynamics during the active interaction of actin and myosin. We conclude that during the active interaction of actin and ATP with myosin, S1A1 is more effective at stabilizing the S state (probably the force-generating state) of actomyosin, while S1A2 tends to stabilize the weak-binding (non-force-generating) W state. When a mixture of isoenzymes is present, S1A1 is dominant in its effects on actin dynamics. We conclude that ELC of skeletal muscle myosin modulates strong-to-weak structural transitions during the actomyosin ATPase cycle in an isoform-dependent manner, with significant implications for the contractile function of actomyosin.
authors: Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455, USA.
authors: Ewa
authors: E
authors: Prochniewicz
authors: Piyali
authors: P
authors: Guhathakurta
authors: David D
authors: DD
authors: Thomas
mesh_terms: D000199:Actins / Q000032:analysis / Q000378:metabolism*; D000205:Actomyosin / Q000032:analysis / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D016880:Anisotropy; D049408:Luminescent Agents / Q000032:analysis; D008163:Luminescent Measurements; D008958:Models, Molecular; D018482:Muscle, Skeletal / Q000737:chemistry / Q000378:metabolism; D018994:Myosin Light Chains / Q000032:analysis / Q000378:metabolism*; D009994:Osmolar Concentration; D020033:Protein Isoforms / Q000032:analysis / Q000378:metabolism; D011817:Rabbits
chemical_list: D000199:Actins; D049408:Luminescent Agents; D018994:Myosin Light Chains; D020033:Protein Isoforms; D000255:Adenosine Triphosphate; D000205:Actomyosin; D000251:Adenosine Triphosphatases