Confirming the 3D Solution Structure of a Short Double-Stranded DNA Sequence Using NMR Spectroscopy

Abstract

Nuclear magnetic resonance (NMR) spectroscopy is an indispensable tool used for the elucidation and confirmation of the solution structure of molecules that was first reported in 1938 by Isidor Rabi (1). A mere eight years later, its use was adapted for structure elucidation of liquids and solids by Felix Bloch and Edward Mills Purcell (2, 3), who four years later shared a Nobel Prize for their work. In more recent times, NMR spectroscopy has been extended to the study of larger biomolecules and has become a technique of paramount importance. In order to introduce students to this field of study, it is imperative that there be a sound understanding of how this technique works. Veeraraghavan describes a short course that effectively introduces students to the basics of NMR spectroscopy (4). A review of the articles available through this Journal shows examples of NMR studies of biological molecules such as peptides, phospholipids, and cytochrome c (5-8); however, there are no examples that use NMR spectroscopy to investigate DNA structure. The work herein represents a 6-h laboratory workshop with the aim of confirming the 3D structure of a short doublestrand DNA sequence, achieved using 2D 1H NOESY NMR spectroscopy, with the aid of a computer-generated model.