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• W83633 abstract "This chapter summarizes the application and advantages of hydroxyl radicals for probing protein-binding sites on Ribonucleic acid (RNAs). Hydroxyl radicals are small, readily diffusible reagents that cleave ribose groups of the RNA backbone without preference for nucleotide or secondary structure character. Because of the small size of the hydroxyl radical reagent, the resulting footprinting data have much higher resolution than that offered by enzymatic cleavage methods. A stepwise approach for preparing the RNA transcripts, radiolabeling, protein binding, Fe(II)-ethylenediaminetetraacetic acid (EDTA) cleavage, and data analysis are provided; moreover, the advantages and experimental considerations relevant to the technique are described. Examples of hydroxyl radical footprinting are drawn from the analysis of RNA–coat protein interactions among the single-stranded, positive-sense RNA viruses, alfalfa mosaic virus, and the ilarviruses. The chapter discusses in detail about the prior uses of hydroxyl radical footprinting. The chapter also explains concepts related to preparing an RNA template for hydroxyl radical footprinting. Concepts of binding and cleavage reactions are also discussed. The chapter also presents an overview of footprinting data, and coupling hydroxyl radical footprinting with other methods." @default.
• W83633 created "2016-06-24" @default.
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• W83633 date "1997-01-01" @default.
• W83633 modified "2023-09-26" @default.
• W83633 title "Footprinting RNA—Protein Complexes with Hydroxyl Radicals" @default.
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• W83633 doi "https://doi.org/10.1016/b978-012587545-5/50016-1" @default.
• W83633 hasPublicationYear "1997" @default.
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