The overall goal of the proposed research is to empower NMR spectroscopy for characterizing large and biologically significant proteins and protein complexes. This will lead to new insights into biological mechanisms and will ultimately lead to improving human health. Proteins function primarily by interacting with other proteins, nucleic acids, small ligands or substrates. Thus, the overall goal of this grant is to understand mechanism based on structural insights, with a focus on protein interactions. This will be achieved with developing a set of NMR methods that can characterize challenging protein complexes in the range of 50 kDa and beyond. All components of the proposal contain both technology developments and applications to important targets that play roles in disease, are potential drug targets, or producers of natural products. Component 1 (Wagner) is on NMR approaches for structural characterization of large proteins and protein complexes. It will develop new methods for studies of proteins and protein complexes and will apply these for characterizing interactions of a viral internal ribosome entry site (IRES) RNA with translation initiation factors. Component 2 (Wagner/Walsh) is on the Enterobactin non-ribosomal peptide synthetase. It will solve structures of large mufti-domain units of this system and determine structures of complexes between EntF and EntB. Component 3 (Reinherz) will elucidate the structure and function of the signaling elements of the T-cell receptor. Component 4 (Naar) will investigate the structure and function of the pleiotropic drug resistance system of pathogenic yeast. The research will be supported by three cores for Administration (Core A), NMR instrumentation and Chemistry (Core B) and Computation (Core C). The interaction between the research components and cores of this POI grant simulated the development of many new NMR and data processing techniques that now facilitate structural studies of challenging macromolecular systems. We anticipate that the proposed research will continue to advance the capabilities of NMR for structural biology.

Public Health Relevance

The research proposed by the closely interacting research components will enhance the capabilities of NMR for solving structures of larger and more complex proteins and protein complexes. Elucidating how proteins interact will open avenues for understanding biological mechanisms and will point to new possibilities for drug design and cure human diseases.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Program Projects (P01)
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Special Emphasis Panel (ZRG1-BCMB-S (40))
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Wehrle, Janna P
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Harvard University
Schools of Medicine
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Takeuchi, Koh; Sun, Zhen-Yu J; Li, Shuai et al. (2015) NMR resonance assignments of the catalytic domain of human serine/threonine phosphatase calcineurin in unligated and PVIVIT-peptide-bound states. Biomol NMR Assign 9:201-5
Edmonds, Katherine A; Wagner, Gerhard (2015) (1)H, (13)C, and (15)N backbone and sidechain chemical shift assignments for the HEAT2 domain of human eIF4GI. Biomol NMR Assign 9:157-60
Hagn, Franz; Wagner, Gerhard (2015) Structure refinement and membrane positioning of selectively labeled OmpX in phospholipid nanodiscs. J Biomol NMR 61:249-60
Wommack, Andrew J; Ziarek, Joshua J; Tomaras, Jill et al. (2014) Discovery and characterization of a disulfide-locked C(2)-symmetric defensin peptide. J Am Chem Soc 136:13494-7
Boeszoermenyi, Andras; Schmidt, Jens C; Cheeseman, Iain M et al. (2014) Resonance assignments of the microtubule-binding domain of the C. elegans spindle and kinetochore-associated protein 1. Biomol NMR Assign 8:275-8
Akabayov, Sabine R; Akabayov, Barak; Wagner, Gerhard (2014) Human translation initiation factor eIF4G1 possesses a low-affinity ATP binding site facing the ATP-binding cleft of eIF4A in the eIF4G/eIF4A complex. Biochemistry 53:6422-5
Sun, Zhen-Yu J; Cheng, Yuxing; Kim, Mikyung et al. (2014) Disruption of helix-capping residues 671 and 674 reveals a role in HIV-1 entry for a specialized hinge segment of the membrane proximal external region of gp41. J Mol Biol 426:1095-108
Hoch, Jeffrey C; Maciejewski, Mark W; Mobli, Mehdi et al. (2014) Nonuniform sampling and maximum entropy reconstruction in multidimensional NMR. Acc Chem Res 47:708-17
Brazin, Kristine N; Mallis, Robert J; Li, Chen et al. (2014) Constitutively oxidized CXXC motifs within the CD3 heterodimeric ectodomains of the T cell receptor complex enforce the conformation of juxtaposed segments. J Biol Chem 289:18880-92
Linser, Rasmus; Bardiaux, Benjamin; Andreas, Loren B et al. (2014) Solid-state NMR structure determination from diagonal-compensated, sparsely nonuniform-sampled 4D proton-proton restraints. J Am Chem Soc 136:11002-10

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