Computer Assisted Assignment of13C or15N Edited 3D-NOESY–HSQC Spectra Using Back Calculated and Experimental Spectra

Abstract A new tool, for the simulation of 15 N or 13 C edited 3D-NOESY–HSQC spectra using the complete relaxation matrix approach, has been developed and integrated in the program AURELIA. This tool should be particularly useful for the fast and reliable computer assisted assignment of 3D-NOESY–HSQC spectra by comparing back-calculated and experimental spectra in an iterative process. Folded spectra are sometimes used to enhance the digital resolution in the indirect dimensions of multidimensional spectra. However, these spectra are usually difficult to analyze. To simplify this assignment process we have incorporated the simulation and automated annotation of folded peaks into the program. It is hereby possible to simulate multiple folding in all three dimensions of 3D 15 N- or 13 C-NOESY–HSQC spectra. By comparing experimental 3D-NOESY–HSQC spectra with spectra back calculated from a single trial structure or a set of trial structures, a user can easily check if the final structures explain all experimental NOEs. The new feature has been successfully tested with the histidine-containing phosphocarrier protein HPr from Staphylococcus carnosus.