Toward direct determination of conformations of protein building units from multidimensional NMR experiments III

Abstract

Chemical shielding anisotropy tensors have been determined, within the GIAO-RHF formalism using a smaller (6-31+G(d)) and two medium-size basis sets (6-311++G(d,p) and TZ2P), for all elements of the conformational library (altogether 27 structures) of the hydrophobic model peptide For{L-Phe{NH2. The individual chemical shifts and their conformational averages have been compared to their experimental counterparts taken from the BioMagnetic Resonance Bank (BMRB). At the highest level of theory applied, for all nuclei but the amide proton, deviations between statistically averaged theoretical and experimental chemical shifts are as low as a few percent. One-dimensional (1D) chemical shift { structure plots do not allow unambiguous identication of backbone conformations. On the other hand, on chemical shift { chemical shift plots of selected nuclei, e.g., 1 H N with 15 No r 15 Nw ith 13 C, regions corresponding to major conformational motifs have been found, providing basis for the identication of peptide conformers solely from NMR shift data. The 2D 1 H{ 13 C as well as the 3D 1 H{ 13 C{ 13 C chemical shift { chemical shift plots appear to be of special importance for direct determination of conformations of protein building units from multidimensional NMR experiments. 48 pairs of 1 H{ 13 C data for phenylalanine residues have been extracted from 18 selected proteins and compared to relevant ab initio results, supporting the calculated results. Thus, the appealing idea of establishing backbone folding information of peptides and proteins from chemical shift information alone, obtained from selected multiple-pulse NMR experiments (e.g., 2D-HSQC, 2D-HMQC, and 3D-HNCA), has received further support.