利用X射线对RNA进行时间分辨的羟基自由基足迹分析
Time-Resolved Hydroxyl Radical Footprinting of RNA with X-Rays.
作者信息
Hao Yumeng, Bohon Jen, Hulscher Ryan, Rappé Mollie C, Gupta Sayan, Adilakshmi Tadepalli, Woodson Sarah A
机构信息
Johns Hopkins University, Baltimore, Maryland.
Center for Synchrotron Biosciences, Case Western Reserve University, Cleveland, Ohio.
出版信息
Curr Protoc Nucleic Acid Chem. 2018 Jun;73(1):e52. doi: 10.1002/cpnc.52. Epub 2018 Jun 7.
Abstract
RNA footprinting by hydroxyl radical cleavage provides 'snapshots' of RNA tertiary structure or protein interactions that bury the RNA backbone. Generation of hydroxyl radicals with a high-flux synchrotron X-ray beam provides analysis on a short timescale (5-100 msec), which enables the structures of folding intermediates or other transient conformational states to be determined in biochemical solutions or cells. This article provides protocols for using synchrotron beamlines for hydroxyl radical footprinting. © 2018 by John Wiley & Sons, Inc.
摘要
通过羟基自由基切割进行的RNA足迹分析提供了RNA三级结构或掩埋RNA主链的蛋白质相互作用的“快照”。利用高通量同步加速器X射线束产生羟基自由基可在短时间尺度（5 - 100毫秒）内进行分析，这使得能够在生化溶液或细胞中确定折叠中间体或其他瞬态构象状态的结构。本文提供了使用同步加速器光束线进行羟基自由基足迹分析的方案。© 2018约翰威立国际出版公司。
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