This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.We are developing HIFI methods to speed up data collection of NMR spectra for the assignments of backbone and side chain resonances. In HIFI multidimensional experiments are collected as two-dimensional tilted planes, where the indirect dimension is a mixture of two or more indirectly evolved dimensions. The tilt angle for the combined indirect dimension is adaptively estimated for each plane in real time to reduce overlap and maximize the amount of information that can be extracted from each experiment. In order to apply our methods to larger proteins, we utilize a new labeling scheme where proteins are produced using cell free technology with SAIL labeled amino acids. In SAIL labeled amino acids some of the protons in the side chain are replaced by deuterium, thus reducing proton density in the protein. SAIL labeling has the advantage of reducing ambiguity in proton assignments while at the same time improving the quality of the spectra by decreasing the speed at which NMR signal are relaxing (fast relaxation is one of the main obstacles in studying large proteins by NMR).Our goal is to speed up the time required for collecting the NMR experiments that are required for assigning signals in NMR spectra, while at the same time extending the limit of proteins that can be studied by to larger molecules.
Showing the most recent 10 out of 613 publications
