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首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Theory of single-molecule controlled rotation experiments predictions tests and comparison with stalling experiments in F1-ATPase
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Theory of single-molecule controlled rotation experiments predictions tests and comparison with stalling experiments in F1-ATPase

机译：单分子控制旋转实验的原理预测测试以及与F1-ATPase中的失速实验的比较

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A recently proposed chemomechanical group transfer theory of rotary biomolecular motors is applied to treat single-molecule controlled rotation experiments. In these experiments, single-molecule fluorescence is used to measure the binding and release rate constants of nucleotides by monitoring the occupancy of binding sites. It is shown how missed events of nucleotide binding and release in these experiments can be corrected using theory, with F1-ATP synthase as an example. The missed events are significant when the reverse rate is very fast. Using the theory the actual rate constants in the controlled rotation experiments and the corrections are predicted from independent data, including other single-molecule rotation and ensemble biochemical experiments. The effective torsional elastic constant is found to depend on the binding/releasing nucleotide, and it is smaller for ADP than for ATP. There is a good agreement, with no adjustable parameters, between the theoretical and experimental results of controlled rotation experiments and stalling experiments, for the range of angles where the data overlap. This agreement is perhaps all the more surprising because it occurs even though the binding and release of fluorescent nucleotides is monitored at single-site occupancy concentrations, whereas the stalling and free rotation experiments have multiple-site occupancy.
机译：最近提出的旋转生物分子马达的化学机械基团转移理论被用于治疗单分子控制的旋转实验。在这些实验中，单分子荧光用于通过监测结合位点的占据来测量核苷酸的结合和释放速率常数。显示了如何使用F1-ATP合酶为例，通过理论校正这些实验中核苷酸结合和释放的遗漏事件。当反向速率非常快时，错过的事件很重要。使用该理论，可以从独立数据（包括其他单分子旋转和整体生化实验）中预测受控旋转实验中的实际速率常数和校正值。发现有效扭转弹性常数取决于结合/释放核苷酸，并且对于ADP而言，其小于ATP。对于数据重叠的角度范围，在受控旋转实验和失速实验的理论和实验结果之间有一个很好的协议，没有可调整的参数。该协议可能更令人惊讶，因为即使在单点占用浓度下监测荧光核苷酸的结合和释放，它也会发生，而停转和自由旋转实验则具有多点占用。

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