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|Author(s):||B. Luy; John P. Marino;|
|Title:||Measurement and Application of 1H-19F Dipolar Couplings in the Structure Determination of 2'-Fluorolabeled RNA|
|Published:||May 01, 2001|
|Abstract:||Residual dipolar couplings can provide powerful restraints for determination and refinement of the solution structure of macromolecules. The application of these couplings in nucleic acid structure elucidation can have an especially dramatic impact since they provide long-range restraints, typically absent in NOE and J-coupling measurements. Here we describe sensitive X-filtered-E.COSY-type methods designed to measure both the sign and magnitude of long-range 1H-19F dipolar couplings on selectively fluorine labeled RNA oligonucleotides oriented in solution by a liquid crystalline medium. The techniques for measuring 1H-19F dipolar couplings are demonstrated on a 21mer RNA hairpin, which has been specifically labeled with fluorine at the 2'-hydroxyl position of three ribose sugars. Experimentally measured 1H-19F dipolar couplings for the 2'-deoxy-2'-fluoro-sugars located in the helical region of the RNA hairpin were found to be in excellent agreement with values predicted using canonical A-form helical geometry, demonstrating that these couplings can provide accurate restraints for the refinement of RNA structures determined by NMR.|
|Citation:||Journal of Biomolecular Nmr|
|Pages:||pp. 39 - 47|
|Research Areas:||DNA, Bioscience & Health, Chemistry, Measurements|