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通过核磁共振光谱法检测溶液中和晶体中的大肠杆菌5S rRNA E结构域的多种构象。

Detection of multiple conformations of the E-domain of 5S rRNA from Escherichia coli in solution and in crystals by NMR spectroscopy.

作者信息

Grüne M, Fürste J P, Klussmann S, Erdmann V A, Brown L R

机构信息

Institut für Molekulare Biotechnologie e.V., Jena, Germany.

出版信息

Nucleic Acids Res. 1996 Jul 1;24(13):2592-6. doi: 10.1093/nar/24.13.2592.

Abstract

NMR spectroscopy of the E-domain fragment of Escherichia coli 5S rRNA indicates that this molecule exists in solution as either a stem-loop or as a duplex with two U-U base pairs in the bulge region. At temperatures below 27 degrees C, interconversion between the monomeric and dimeric forms in solution occurs on a time scale of weeks and allows the preparation of samples on which NMR structure determinations can be carried out on predominantly monomeric or dimeric species. The NMR results obtained provide comparison data for the distinction between A- and B-form E.coli 5S rRNA and for the possible kinetics of conversion between these forms. NMR evidence is presented that the duplex form also exists in crystals and suggestions are made for means to obtain stem-loop conformations of E-domain and other small RNA stem-loop sequences in crystals.

摘要

大肠杆菌5S rRNA E结构域片段的核磁共振光谱表明,该分子在溶液中以茎环结构或在凸起区域具有两个U-U碱基对的双链体形式存在。在低于27摄氏度的温度下,溶液中单体和二聚体形式之间的相互转化在数周的时间尺度上发生,这使得能够制备样品,在这些样品上可以对主要为单体或二聚体的物种进行核磁共振结构测定。所获得的核磁共振结果为区分A型和B型大肠杆菌5S rRNA以及这些形式之间可能的转化动力学提供了比较数据。有核磁共振证据表明双链体形式也存在于晶体中,并提出了在晶体中获得E结构域和其他小RNA茎环序列的茎环构象的方法建议。

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