Crystallographic studies of RNA/protein complexes are primordial for the understanding of recognition determinants and catalytic mechanisms in the case of enzymes. However, due to the flexibility and propensity to conformational heterogeneity of RNAs, as well as the mostly electrostatic interactions of RNA/protein complexes, they are difficult to crystallize. We present here a method to trap the two interacting partners in a covalent complex, based on a modified reactive RNA allowing the use of the full range of common crystallogenesis tools. We demonstrate the practicability of our approach with the production of a covalent complex of the Thermus thermophilus m1A58 tRNA modification enzyme, and a modified stem loop mimicking the natural substrate of the enzyme.

中文翻译：

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用于结构研究的交联RNA /蛋白质复合物的设计。

RNA /蛋白质复合物的晶体学研究对于理解酶的识别决定簇和催化机制是至关重要的。然而，由于RNA的构象异质性的灵活性和倾向性，以及RNA /蛋白质复合物的大部分静电相互作用，它们难以结晶。我们在此介绍一种基于修饰的反应性RNA来捕获共价复合物中的两个相互作用伙伴的方法，该方法允许使用所有范围的常见结晶生成工具。我们证明了我们的方法的实用性与嗜热菌Thermus m1A58 tRNA修饰酶的共价复合物的生产，以及模拟该酶天然底物的修饰茎环。