Use of RNA-bound Tb3+ as a FRET donor

Lanthanide ions such as Tb3+ and Eu3+ have long been used to probe RNA and protein structures due to their luminescence properties and their steric and chemical similarities to biological metal ions such as Mg2+ and Ca2+. In this article, we introduce a method that utilizes the enhanced Tb3+ lumines...

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Bibliographic Details
Published in:Methods (San Diego, Calif.) Calif.), 2010-10, Vol.52 (2), p.173-179
Main Authors: Yuan, Faqing, Greenbaum, Nancy L.
Format: Article
Language:English
Subjects:
Fluorescence
Fluorescence Resonance Energy Transfer - methods
FRET
Luminescence
Nucleic Acid Conformation
RNA
RNA - chemistry
Tb3
Terbium - analysis
Terbium - chemistry
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Summary:Lanthanide ions such as Tb3+ and Eu3+ have long been used to probe RNA and protein structures due to their luminescence properties and their steric and chemical similarities to biological metal ions such as Mg2+ and Ca2+. In this article, we introduce a method that utilizes the enhanced Tb3+ luminescence upon site-binding to RNA molecules as a FRET donor. Using this method, it is possible to identify specific metal ion-binding locations within a folded RNA molecule. Applications of this method include introducing FRET donors into molecules of interest by engineering RNA loops or bulges as Tb3+-binding pockets.
ISSN:1046-2023
1095-9130
DOI:10.1016/j.ymeth.2010.08.008