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BCMBJHU SOM

Faculty & Research

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Dominique Frueh

Department Affiliation Primary: Biophysics & Biophysical Chemistry
RankAssistant Professor
Phone NumbersOffice: 410-614-4719
Lab: 410-614-2229
Fax: 410-955-0637
Emaildfrueh1@jhmi.edu
School of Medicine Address725 N. Wolfe Street
701 Hunterian
Baltimore MD 21205
Link to Lab Homepage
Dominique Frueh

Research Topic: Structural and dynamic studies of active enzymatic systems by Nuclear Magnetic Resonance (NMR) - Mechanisms of domain communication in Non-ribosomal Peptide Synthetases.

Our laboratory uses nuclear magnetic resonance (NMR) to study modulations of protein dynamics and conformations in active enzymatic systems.

Non-ribosomal peptide synthetases (NRPSs) are large enzymatic systems responsible for the biosynthesis of a wealth of secondary metabolites, many of which are used by pharmaceutical scientists to produce drugs such as antibiotics or anticancer agents. To synthesize all of these remarkably diverse compounds, bacteria and fungi use a surprisingly conserved strategy: NRPSs are organized in modules, made of conserved domains, that each incorporates a dedicated substrate. Thus, new compounds with improved activities can be generated, in principle, by swapping domains or modules in order to control substrate incorporation and hence the final product. To do this efficiently, a deep understanding of domain communication during the synthesis is required. NMR and crystallography studies indicate that domain interactions are transient and that their quaternary structure is likely subject to rearrangements during the synthesis. In addition, both techniques show that individual domains are subject to conformational heterogeneity. We principally use NMR to investigate inter- and intra-domain modifications occurring during the catalytic steps of non-ribosomal peptide synthesis.

Development of NMR methods. The enzymatic systems that we investigate provide many challenges to NMR. Some of the domains or multi-domains are large, and therefore, the resulting data suffer from spectral crowding and signal losses. In addition, the proteins are subject to dynamics, which may further deteriorate the quality of the spectra.Consequently, new methods often need to be developed to overcome these challenges. The techniques provide means to assign the NMR signals to the atoms in the proteins, to measure structural constraints, or to monitor dynamics. When faced with an impasse, new techniques can be designed in our lab. With these new techniques available, new protein targets can be considered, thereby increasing our overall understanding of the systems. Thus, we foster a synergy between studies of relevant biological systems and the development of new NMR techniques.

Publications:

Frueh DP Arthanari H, Koglin A, Walsh CT, Wagner G. A double TROSY hNCAnH experiment for efficient assignment of large and challenging proteins. J Am Chem Soc. 2009 Sep 16;131(36):12880-1.
PubMed
 

Frueh DP, Arthanari H, Koglin A, Vosburg DA, Bennett AE, Walsh CT, Wagner G. Dynamic thiolation-thioesterase structure of a non-ribosomal peptide synthetase. Nature. 2008 Aug 14;454(7206):903-6.
PubMed

Selenko, P., Frueh DP, Elsaesser S, Haas W, Gygi SP, Ruderman J, Wagner G, In situ observation of protein phosphorylation by high-resolution NMR spectroscopy. Nat Struct Mol Biol. 2008, 15(3):321-9.
PubMed

Frueh D, Internal Motions in Proteins and Interference Effects in Nuclear Magnetic Resonance, Prog. NMR spectrosc., 2002, 41, 305-324.
Link

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