Sequence-specific solution structure of d-GGTACGCGTACC

Abstract

Complete resonance assignments of nonexchangeable base protons and sugar protons in d-GGTACGCGTACC at 500 MHz have been obtained by two-dimensional correlated spectroscopy (COSY) and nuclear Overhauser enhancement spectroscopy (NOESY). The characteristic phase-sensitive multiplet patterns of the intrasugar cross peaks in the .omega.1-scaled COSY spectrum have been used to estimate several scalar coupling constants (J). These coupling constants combined with the intranucleotide COSY cross peak intensities have been used to identify the sugar pucker of individual nucleotide units. In most cases, the deoxyribose rings adopt a conformation close to O4''-endo. Spin-diffusion has been monitored from the buildup of the normalized volumes of NOE cross peaks in NOESY spectra as a function of mixing time. A set of 55 intranucleotide and internucleotide interproton distances have been estimated from the low mixing time NOESY spectrum (.tau.m = 75 ms). The estimated intranucleotide proton-proton distances have been used to determine the individual glycoside dihedral angles of the nucleotide units which lie in the anti domain. It is observed that the molecule adopts an overall confirmation close to that of the B-form although there are differences in the intricate details.