This research component has the overall goal to develop and improve methods for characterizing large proteins and protein complexes. Protein interactions are crucial for many biological processes, such as cellular switches, signaling mechanisms, enzyme regulation, gene expression and development. Much is known 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 in contrast to the fact that many protein interactions must be weak and transient, for example to rapidly turn signaling pathways on and off, or to recycle cellular proteins. Furthermore, protein complexes represent an underutilized class of targets for design of drugs against diseases. NMR has unique capabilities for defining structures and states of large proteins and protein complexes that cannot be obtained with crystallography. Major improvements of NMR hardware have recently become available that are only beginning to be efficiently utilized. New ideas of spin physics, control theory, sampling strategies, signal processing, or data analysis are being developed, together with new strategies for sample preparation. All this promises major advances of NMR spectroscopy with large proteins and protein complexes. Here we propose to develop new approaches to make optimal use of modern NMR hardware to gain new insights into structures of large proteins and protein complexes. This will require abandoning some of the traditional procedures for data acquisition and require new data processing methods. This grant has spearheaded 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 experiments proposed employ heavily non-uniform sampling methods, coherence co-evolution procedures, and they are geared towards optimum use of high-field instruments and use optimum control theory for pulse sequence design.
Aim 2. Approaches for characterizing protein complexes. This includes new co-expression methods for facilitating NMR studies of complexes, development of new protein tags to improve the solution behavior of complexes, and NMR and computational techniques for defining the arrangement of proteins in tight and in weak complexes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM047467-19
Application #
8072536
Study Section
Special Emphasis Panel (ZRG1)
Project Start
2010-05-01
Project End
2012-04-30
Budget Start
2010-05-01
Budget End
2011-04-30
Support Year
19
Fiscal Year
2010
Total Cost
$265,815
Indirect Cost
Name
Harvard University
Department
Type
DUNS #
047006379
City
Boston
State
MA
Country
United States
Zip Code
02115
Brazin, Kristine N; Mallis, Robert J; Boeszoermenyi, Andras et al. (2018) The T Cell Antigen Receptor ? Transmembrane Domain Coordinates Triggering through Regulation of Bilayer Immersion and CD3 Subunit Associations. Immunity 49:829-841.e6
Chhabra, Sandeep; Fischer, Patrick; Takeuchi, Koh et al. (2018) 15N detection harnesses the slow relaxation property of nitrogen: Delivering enhanced resolution for intrinsically disordered proteins. Proc Natl Acad Sci U S A 115:E1710-E1719
Zhao, Zhao; Zhang, Meng; Hogle, James M et al. (2018) DNA-Corralled Nanodiscs for the Structural and Functional Characterization of Membrane Proteins and Viral Entry. J Am Chem Soc 140:10639-10643
Hagn, Franz; Nasr, Mahmoud L; Wagner, Gerhard (2018) Assembly of phospholipid nanodiscs of controlled size for structural studies of membrane proteins by NMR. Nat Protoc 13:79-98
Nasr, Mahmoud L; Wagner, Gerhard (2018) Covalently circularized nanodiscs; challenges and applications. Curr Opin Struct Biol 51:129-134
Coote, Paul W; Robson, Scott A; Dubey, Abhinav et al. (2018) Optimal control theory enables homonuclear decoupling without Bloch-Siegert shifts in NMR spectroscopy. Nat Commun 9:3014
Ziarek, Joshua J; Baptista, Diego; Wagner, Gerhard (2018) Recent developments in solution nuclear magnetic resonance (NMR)-based molecular biology. J Mol Med (Berl) 96:1-8
Näär, Anders M (2018) miR-33: A Metabolic Conundrum. Trends Endocrinol Metab 29:667-668
Nasr, Mahmoud L; Baptista, Diego; Strauss, Mike et al. (2017) Covalently circularized nanodiscs for studying membrane proteins and viral entry. Nat Methods 14:49-52
Feng, Yinnian; Brazin, Kristine N; Kobayashi, Eiji et al. (2017) Mechanosensing drives acuity of ?? T-cell recognition. Proc Natl Acad Sci U S A 114:E8204-E8213

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