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The model fit also supplies an estimate of the unloaded rate of rear-head release, 260 ± 10 s−10
The fit parameters indicate that WT levels of inter-head tension can amplify the rate of rear-head release by an order of magnitude beyond its unloaded rate.
However, because this unloaded rate is already more than double the rate of the hydrolysis-associated events of the kinesin cycle, k2, the rate-determining step of the full cycle cannot be rear-head release.
Measurements of unbinding rates under load were fit to the exponential function, k off = k off 0 exp [ | F trap | δ off / k B T ] where k off 0 is the unloaded rate and δoff is the associated distance parameter.
From the 2-HB state [3], the final transition of the cycle consists of rear-head release (which is also load-dependent) and is modeled as k 3 = k 3 0 exp [ (F trap + F i ) δ 3 / k B T ], where k 3 0 is the unloaded rate constant and δ3 is the associated distance parameter.
The first transition in this cycle consists of the force-producing mechanical step, [1] → [2], which is modeled by a force-dependent rate, k 1 = k 1 0 exp [ F trap δ 1 / k B T ] where k 1 0 is the unloaded rate constant, Ftrap is the applied force, δ1 is a characteristic distance parameter, and kB T is Boltzmann's constant times the absolute temperature.
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Here, dstep is the kinesin step size (fixed at 8.2 nm), Fi is inter-head tension, kB T is Boltzmann's constant times the absolute temperature, k n 0 are the unloaded rates of the 3-state model, and δ n are the corresponding characteristic distance parameters for these rates.
The unloaded release rate (k off) and the associated distance parameter (δoff) were obtained from exponential fits to unbinding data acquired under hindering loads, −25 to 0 pN, and assisting loads, +2 to +20 pN.
Following two minutes of rest and one minute of unloaded cycling, work rate was increased every minute by a predetermined amount, from 5 to 20 W.min-1 according to the participant's self-reported exercise capacity and disease severity so that the test was approximately 10 minutes duration [ 21].
The same dwells must also limit the rate of unloaded hydrolysis.
It is a multiple unitary step motor characteristic proportional to the rate of unloaded cross-bridge cycling with the higher rate producing a more frequent jth step.
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