Abstract

The overall goal of this project is to learn about the interaction of domains in non-ribosomal polypeptide synthetase modules, using the 142 kDa E. coli EntF. This protein consists of four domains, C, A, T and TE that act as chain elongation and termination module to make and release the enterobactin siderophore. We are interested in how the domains cooperate to synthesize enterobactin, and how the HS-pantetheinyl-P group bound to the TE domain affects the interaction of domains, and how this may relate to the assembly- line function of the protein. The multidomain protein is a model system for development of NMR for large proteins The specific aims are:
Specific Aim 1.. Complete structure determination of the 36 kDa A-T didomain fragment of the 142kDa EntF Specific Aim 2 Studies of structures and interactions of the in cis C and A domains centering on the T domain scaffold and how these partner proteins dock with it SpecificAim 3. Studies of the structure of the A-T 70 kDa didomain of EntF Specific Aim 4. . Interaction of the HS-pantetheinyl-P prosthetic group (isotopically labeled or spin-labeled )on the two T domains in EntB and EntF with different domains in cis and in trans (C, A, TE) in unacylated and acylated forms

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM047467-17
Application #
7618427
Study Section
Special Emphasis Panel (ZRG1)
Project Start
Project End
Budget Start
2008-05-01
Budget End
2009-04-30
Support Year
17
Fiscal Year
2008
Total Cost
$399,905
Indirect Cost

  Institution

Name
Harvard University
Department
Type
DUNS #
047006379
City
Boston
State
MA
Country
United States
Zip Code
02115

  Publications

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
Hyberts, Sven G; Robson, Scott A; Wagner, Gerhard (2017) Interpolating and extrapolating with hmsIST: seeking a tmax for optimal sensitivity, resolution and frequency accuracy. J Biomol NMR 68:139-154
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

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