471 research outputs found
Supplemental Material for Xu and Yanagida, 2019
No full textTable S1 and S2 contain lists the primers used to construct cut14 and cut3 mutants, respectively. Figure S1 describes PCR conditions used to introduce mutations into target genes. Figure S2 describes transformation protocol
The synergic role of actomyosin architecture and biased detachment in muscle energetics: Insights in cross bridge mechanism beyond the lever‐arm swing
No full textMuscle energetics reflects the ability of myosin motors to convert chemical energy into mechanical energy. How this process takes place remains one of the most elusive questions in the field. Here, we combined experimental measurements of in vitro sliding velocity based on DNA-origami built filaments carrying myosins with different lever arm length and Monte Carlo simulations based on a model which accounts for three basic components: (i) the geometrical hindrance, (ii) the mechano‐sensing mechanism, and (iii) the biased kinetics for stretched or compressed motors. The model simulations showed that the geometrical hindrance due to acto-myosin spatial mismatching and the preferential detachment of compressed motors are synergic in generating the rapid increase in the ATP‐ase rate from isometric to moderate velocities of contraction, thus acting as an energy‐conservation strategy in muscle contraction. The velocity measurements on a DNA‐origami filament that preserves the motors’ distribution showed that geometrical hindrance and biased detachment generate a non‐zero sliding velocity even without rotation of the myosin lever‐arm, which is widely recognized as the basic event in muscle contraction. Because biased detachment is a mechanism for the rectification of thermal fluctuations, in the Brownian‐ratchet framework, we predict that it requires a non‐negligible amount of energy to preserve the second law of thermodynamics. Taken together, our theoretical and experimental results elucidate less considered components in the chemo‐mechanical energy transduction in muscle
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