Measurement and Application of 1H-19F Dipolar Couplings in the Structure Determination of 2'-Fluorolabeled RNA

Published

May 1, 2001

Author(s)

B. Luy, John Marino

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

Volume

20

Issue

No. 1

Pub Type

Journals

Keywords

dipolar couplings, dipolar-TOCSY, E.COSY, fluorine, MOCCA, RNA

Citation

Luy, B. and Marino, J. (2001), Measurement and Application of ¹H-¹⁹F Dipolar Couplings in the Structure Determination of 2'-Fluorolabeled RNA, Journal of Biomolecular Nmr (Accessed March 6, 2025)