The Bridge Helix of RNA polymerase acts as a central nanomechanical switchboard for coordinating catalysis and substrate movement.

The Bridge Helix of RNA polymerase acts as a central nanomechanical switchboard for coordinating catalysis and substrate movement.
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DOI:
10.1155/2011/608385
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发表时间:
2011
期刊:
Archaea (Vancouver, B.C.)
影响因子:
--
通讯作者:
Weinzierl RO
Weinzierl RO
中科院分区:
其他
文献类型:
--
作者:
Weinzierl RO

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在体外组装系统的可用性,以产生重组古细菌RNA聚合酶(RNAP)提供了一个最强大的实验工具,调查仍然相对知之甚少的RNAP功能的分子机制。在过去的几年里,我们开创了新的基于机器人的高通量诱变方法,以研究催化中心周围各种结构域内的结构/功能关系。桥受阻结构域在许多X射线结构中以35个氨基酸长的α螺旋形式出现,在催化过程中通过两个离散分子铰链的扭结协调其他几个结构域的协同运动。影响这些扭结机制的突变对RNAP的特定催化活性有直接影响,在某些情况下可以使其增加一倍以上。分子动力学模拟已经确立了自己作为非常有用的提供额外的见解和详细的模型来解释潜在的结构运动。
The availability of in vitro assembly systems to produce recombinant archaeal RNA polymerases (RNAPs) offers one of the most powerful experimental tools for investigating the still relatively poorly understood molecular mechanisms underlying RNAP function. Over the last few years, we pioneered new robot-based high-throughput mutagenesis approaches to study structure/function relationships within various domains surrounding the catalytic center. The Bridge Helix domain, which appears in numerous X-ray structures as a 35-amino-acid-long alpha helix, coordinates the concerted movement of several other domains during catalysis through kinking of two discrete molecular hinges. Mutations affecting these kinking mechanisms have a direct effect on the specific catalytic activity of RNAP and can in some instances more than double it. Molecular dynamics simulations have established themselves as exceptionally useful for providing additional insights and detailed models to explain the underlying structural motions.
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