This research component has the overall goal to develop and improve methods for characterizing largeproteins and protein complexes. Protein interactions are crucial for many biological processes, such ascellular switches, signaling mechanisms, enzyme regulation, gene expression and development. Much isknown about structures of tight complexes from crystallography and NMR spectroscopy. However,information on weak complexes is rather sparse due to the limitations of current technologies. This is incontrast to the fact that many protein interactions must be weak and transient, for example to rapidly turnsignaling pathways on and off, or to recycle cellular proteins. Furthermore, protein complexes represent anunderutilized class of targets for design of drugs against diseases. NMR has unique capabilities for defining structures and states of large proteins and protein complexesthat cannot be obtained with crystallography. Major improvements of NMR hardware have recently becomeavailable that are only beginning to be efficiently utilized. New ideas of spin physics, control theory, samplingstrategies, signal processing, or data analysis are being developed, together with new strategies for samplepreparation. All this promises major advances of NMR spectroscopy with large proteins and proteincomplexes. Here we propose to develop new approaches to make optimal use of modern NMR hardware to gain newinsights into structures of large proteins and protein complexes. This will require abandoning some of thetraditional procedures for data acquisition and require new data processing methods. This grant hasspearheaded such approaches in the past, and similar efforts have appeared in several other laboratories.We propose research towards two specific aims:
Aim 1. Develop new NMR experiments for characterization of large proteins systems. The experimentsproposed employ heavily non-uniform sampling methods, coherence co-evolution procedures, and they aregeared towards optimum use of high-field instruments and use optimum control theory for pulse sequencedesign.
Aim 2. Approaches for characterizing protein complexes. This includes new co-expression methods forfacilitating NMR studies of complexes, development of new protein tags to improve the solution behavior ofcomplexes, and NMR and computational techniques for defining the arrangement of proteins in tight and inweak complexes.
Showing the most recent 10 out of 245 publications
