John J. Kelley III, Ph.D.
Research Assistant Professor
Donald Lab
004 Sudikoff
Computer Science Department
Dartmouth
Hanover, NH 03755
tel: 603/646-0375

 

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Jack Kelley's research interests are in the fields of Computational Biology and Chemistry. Specifically, developing and implementing experimental planning algorithms for use in high throughput resonance assignment procedures of biologically important molecules. Developing planning algorithms for dual amino acid-selective and site-directed NMR-active labeling of DNA and proteins. Implementing a automated and batch approach of NMR data to perform structure and function calculations on biomolecules. This idea would be to use a minimal set of simple NMR experiments to arrive at structural, dynamical and functional information. In addition, the development of computer- drug design algorithms that can be coupled to or used interactively with structural activity relationships resolved thru NMR. Finally, Jack is interested in pursuing molecular modeling and structural characterization of metal-dna interactions using high field heteronuclear NMR relaxation measurements, coupling constant and distance constraints as input parameters. All areas of his research require an intensive computational component, a chemical physics component and a biological component relating to specific types of cancer.

Publications

  1. John J. Kelley III, Thomas M. Caputo,Steven F. Eaton,Thomas M. Laue & John H. Bushweller. (1997) "Comparison of Backbone Dynamics of Reduced and Oxidized E. coli Glutaredoxin-1 using 15N NMR Relaxation Measurements."Biochemistry 36, 5029-5044.

  2. Xuemei Huang, Barbara E. Crute, Chaohong Sun, Yen-Yee Tang, John J. Kelley III , Amy F. Lewis, Kari L. Hartman, Thomas M. Laue, Nancy A. Speck, & John H. Bushweller. (1998) "Overexpression, Purification, and Biophysical Characterization of the Heterodimerization Domain of the Core-Binding actor b subunit."Journal of Biological Chemistry, 273, 4, 2480-2487

  3. John J. Kelley III & John H. Bushweller. (1998) "1H, 13C and 15N NMR Resonance Assignments of Vaccinia Glutaredoxin-1 in the Fully Reduced Form" Journal of Biomolecular NMR 12, 353-355.

  4. Xian Chen, S.V. Santhana Mariappan, John J. Kelley III , John H. Bushweller, E. Morton Bradbury, & Goutam Gupta. (1998) "A PCR-Based Method for Large Scale Synthesis of Uniformly 13C/15N-Labeled DNA Duplexes." FEBS Letter , in press.

  5. Wang Q., Stacy T., Miller J., Huang X., Kelley J., Binder M., Marin-Padilla M., Sharpe A., Bushweller J., & Speck N. (1997) "Cooperation between the CBF-RD and CBFb subunits of Core Binding Factor" Leukemia (Basingstoke), v.11, no. 12, 2234-2240.





    Previous Research

    Glutaredoxins have been shown to play a number of roles in cells including the reduction of dehydroascorbate, regeneration of oxidatively damaged roteins, acceleration of protein folding, and as a glutathione-dependent source of reducing equivalents for ribonucleotide reductase. The glutaredoxin family of proteins all contain a redox active disulfide in the active site with the sequence -C-P-Y(F)-C- which cycles between the reduced and oxidized forms.
    NMR-based structure determination of E. coli glutaredoxin in its reduced and oxidized forms revealed only subtle structural differences between the two forms. In an effort to characterize the role dynamics may play in the functioning of the protein, we have begun extensive collection of 15N relaxation parameters for E. coli glutaredoxin in several different functional forms. The availability of several different functional forms of the protein including reduced, oxidized, and glutathione mixed disulfide proteins, provides a unique opportunity to examine changes in the dynamics as glutaredoxin cycles through its enzymatic life-cycle.
    In a separate project related to the glutaredoxin family, I have made progress toward the sequence specific backbone resonance assignment of Vaccinia Glutaredoxin. Vaccinia virus is a notorious member of the large DNA viruses called poxviruses which infect vertebrate and invertebrate hosts. The virus encodes many of its own transcriptional enzymes and has the ability to replicate within the cytoplasmic compartment of the cell. Vaccinia Grx is a functional glutaredoxin with thioltransferase and dehydroascorbate reductase activity. Vaccinia Grx contains a biologically important redox-active disulfide bond and is expressed after the onset of DNA replication. Expression of the protein in a bacterial system has allowed us to prepare a homogenous 15N-labelled sample. A number of 3D 15N edited spectra have been recorded including gradient sensitivity enhanced 3D 15N-edited TOCSY, 3D15N-edited NOESY and 2D 15N -edited HSQC to facilitate the assignment of the backbone resonances.