Crossbridge Rotation in EDC-Crosslinked Striated Muscle Fibers
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The rotatability of the strong- and weak-binding myosin heads was tested by stretching glycerinated rabbit psoas fibers after crosslinking the heads to actin by using a carbodiimide EDC. The equatorial 1,1 reflection intensity (I1,1) decreased by ∼10% upon 1% stretch in the presence of various ligands (ATP, ATP-γ-S, pyrophosphate and AMPPNP). As the action of ligands to dissociate actomyosin increased, the relaxation of tension response to stretch and the I1,1 decrease were accelerated. This result is best explained if the ligand converts the crosslinked head to a weak-binding state, in which the head is rotatable because of its acquired elasticity.
Conversely, the weak-to-strong transition was induced in the crosslinked system by removing a ligand (ATP-γ-S) from myosin. Force was produced upon weak-to-strong transition and was accounted for by the increased stiffness of each crosslinked myosin head. However, the comparison of stress-strain curves for the weak- and strong-binding myosin showed that the equilibrium angle of myosin attachment was unchanged, making it unlikely that the weak-to-strong transition is the sole mechanism for active contraction. The calcium-activated force of the same crosslinked fibers showed several features in marked contrast to the force produced by the weak-to-strong transition. This leads to a possibility that the active force is supported by a third class of intermediate which is distinct not only from the weak-binding but also from the strong-binding intermediates in a classical sense.
KeywordsFiber Bundle Fiber Length Active Force Myosin Head Tension Response
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