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. In order to develop experiments for larger proteins, we have successfully prepared a triple labeled sample of Maltodextrin Binding Protein (~43KDa). Our goal is to optimize the parameters and debug existing pulse sequences, as well as, develop new experiments for extending the size of proteins that can be studied by NMR. Some of the methods being considered are the evaluation of the various TROSY techniques, including fast versus sensitivity enhanced experiments, the use of NOESY data to help with resonance assignments for larger proteins, the extension of reduced dimensionality techniques to larger proteins in order to alleviate problems with resonance overlap, the use of parallel evolution to increase the sensitivity of mutidimensional experiments for large proteins characterized by short relaxation delays. We expect our studies to increase our ability for studying larger proteins by NMR and we plan to make our knowledge available to the NMR community.
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