Use of Site-Specifically Tethered Chemical Nucleases to Study Macromolecular Reactions.
Use of Site-Specifically Tethered Chemical Nucleases to Study Macromolecular Reactions.
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使用特定特异性系绳化学核酸酶研究大分子反应。
DOI：
10.1251/bpo49
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发表时间：
2003
影响因子：
6.4
通讯作者：
Sousa R
中科院分区：
文献类型：
--
作者：
Mukherjee S;Sousa R
在复杂的大分子反应过程中，分子构象和与配体的相互作用可能发生多重变化。X射线晶体学只能提供这些变化的有限快照。溶液方法可以增加这样的结构信息，以提供更完整的大分子反应的图片。我们分析了蛋白质构象和蛋白质：核酸相互作用的变化，在转录起始过程中发生的使用化学核酸酶拴到半胱氨酸引入到噬菌体T7（T7 RNAP）的RNA聚合酶的位点特异性。随着聚合酶在转录过程中的步骤，裂解模式的变化揭示了一系列介导转录起始的结构转变。被束缚的化学核酸酶的切割被认为是一种强有力的方法，用于揭示大分子反应的构象动力学，并具有一定的优势，交联或能量转移的方法。
During a complex macromolecular reaction multiple changes in molecular conformation and interactions with ligands may occur. X-ray crystallography may provide only a limited set of snapshots of these changes. Solution methods can augment such structural information to provide a more complete picture of a macromolecular reaction. We analyzed the changes in protein conformation and protein:nucleic acid interactions which occur during transcription initiation by using a chemical nuclease tethered to cysteines introduced site-specifically into the RNA polymerase of bacteriophage T7 (T7 RNAP). Changes in cleavage patterns as the polymerase steps through transcription reveal a series of structural transitions which mediate transcription initiation. Cleavage by tethered chemical nucleases is seen to be a powerful method for revealing the conformational dynamics of macromolecular reactions, and has certain advantages over cross-linking or energy transfer approaches.