Determination of Internucleotide hJ HN Couplings by the Modified 2D J NN-Correlated [ 15N, 1H] TROSY

Abstract

Recent developments in the direct observation of J couplings across hydrogen bonds in proteins and nucleic acids provide additional information for structure and function studies of these molecules by NMR spectroscopy. A J NN-correlated [ 15N, 1H] TROSY experiment proposed by Pervushin et al. (Proc. Natl. Acad. Sci. USA 95, 14147-14151, 1998) can be applied to measure hJ HN in smaller nucleic acids in an E.COSY manner. However, it cannot be effectively applied to large nucleic acids, such as tRNA Trp, since one of the peaks corresponding to a fast relaxing component will be too weak to be observed in the spectra of large molecules. In this Communication, we proposed a modified J NN-correlated [ 15N, 1H] TROSY experiment which enables direct measurement of hJ HN in large nucleic acids. © 2000 Academic Press.