Myosin cleft closure determines the energetics of the actomyosin interaction.
Formation of the strong binding interaction between actin and myosin is essential for force generation in muscle and in cytoskeletal motor systems. To clarify the role of the closure of myosin's actin-binding cleft in the actomyosin interaction, we performed rapid kinetic, spectroscopic, and calorimetric experiments and atomic-level energetic calculations on a variety of myosin isoforms for which atomic structures are available. Surprisingly, we found that the endothermic actin-binding profile of vertebrate skeletal muscle myosin subfragment-1 is unique among studied myosins. We show that the diverse propensity of myosins for cleft closure determines different energetic profiles as well as structural and kinetic pathways of actin binding. Depending on the type of myosin, strong actin binding may occur via induced-fit or conformational preselection mechanisms. However, cleft closure does not directly determine the kinetics and affinity of actin binding. We also show that cleft closure is enthalpically unfavorable, reflecting the development of an internal strain within myosin in order to adopt precise steric complementarity to the actin filament. We propose that cleft closure leads to an increase in the torsional strain of myosin's central β-sheet that has been proposed to serve as an allosteric energy-transducing spring during force generation.
Takács
Balázs
B
Department of Biochemistry, Eötvös University, Pázmány Péter sétány 1/c, H-1117 Budapest, Hungary.
O'Neall-Hennessey
Elizabeth
E
Hetényi
Csaba
C
Kardos
József
J
Szent-Györgyi
Andrew G
AG
Kovács
Mihály
M
D000199:Actins / Q000737:chemistry / Q000378:metabolism; D000205:Actomyosin / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002151:Calorimetry; D007700:Kinetics; D018482:Muscle, Skeletal / Q000378:metabolism*; D015879:Myosin Subfragments / Q000737:chemistry* / Q000378:metabolism; D009218:Myosins / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D013696:Temperature
D000199:Actins; D015879:Myosin Subfragments; D000205:Actomyosin; D009218:Myosins