Protein NMR, structural genomics, proteomics, structural bioinformatics.
For small proteins (<25 kD), distance-geometry
calculations with NMR-derived distance constraints already provide
reliable, high resolution structures. Recent experience indicates
that the same methods can be extended to proteins with molecular
weights up to ca. 40 kD. Approximately 25% of the laboratory effort
is directed toward developing new NMR pulse sequences for solving
larger proteins and for determining protein structures more precisely.
The major part of this effort involves heteronuclear 2D-, 3D-, and
4D- NMR experiments in which magnetization is transferred back and
forth between 1H and 13C
and/or 15N nuclei. These experiments
are carried out with proteins biosynthetically enriched with 13C,
15N, or 2H.
They provide conformation-dependent NMR parameters, which in turn
facilitate protein structure refinement and studies of protein dynamics.
Three- or four-dimensional NMR experiments, using additional 13C
or 15N frequency axes, constitute
an important new approach for unraveling complex spectra of larger
proteins.
The second 25% of the group's work is focused on developing and
implementing computational methods for determining and refining
protein structures based on NMR data. The researchers develop methods
for refining protein structures by comparing experimental and simulated
NMR spectra. Borrowing from the extensive experience of workers
in the energy-refinement field, they utilize molecular-dynamics
and Monte Carlo sampling procedures for overcoming local minima
in distance geometry calculations. Additionally, Montelione's group
develops artificial intelligence computer software for automated
analysis of NMR spectra.
The balance of the laboratory effort centers on determining 3D protein
structures and protein folding. The proteins currently under study
include: growth factors, immunoglobulin-binding proteins, ribonucleases
and both RNA- and DNA-binding proteins. Solution structure analysis
of these proteins will promote a more complete understanding of
their structure-function relationships. A significant portion of
the laboratory effort focuses on applying these methods in the emerging
area of structural genomics (see www.nesg.org).
 |
| Solution NMR structure
of the RNS-binding domain of Non-Structural Protein 1 (NS1)
from Influenza virus; an important target for antiviral drug
design. |
Selected Publications1
Snyder DA, Chen Y, Denissova NG, Acton T, Aramini JM, Ciano M, Karlin R, Liu J, Manor P, Paranji R, Rossi P, Swapna GVT, Xiao R, Rost B, Hunt J, Montelione GT. (2005) Comparisons of NMR spectral quality and success in crystallization demonstrate that NMR and X-ray crystallography are complementary methods for small protein structure determination. J Amer Chem Soc 127:16505-11
Huang YJ, Powers R, Montelione GT.
(2005) Protein NMR recall, precision, and F-measure scores (RPF Scores): Structure quality assessment measures based on information retrieval statistics. J Amer Chem Soc 127:1665-74
Das K, Acton T, Chiang Y, Shih L, Arnold E, Montelione GT. (2004) Crystal structure of RlmAI: Implications for understanding the 23S rRNA G745/G748-methylation at the macrolide antibiotic-binding site. Proc Natl Acad Sci USA 101:4041-6
Montelione GT. (2001) Structural genomics: an approach to the protein folding problem. Proc Natl Acad Sci USA 98:13488-9
Montelione GT, Zheng D, Huang YJ, Gunsalus KC, Szyperski T. (2000) Protein NMR spectroscopy in structural genomics. Nat Struct Biol 7:982-5
Montelione GT, Anderson S.
(1999) Structural genomics: keystone for a Human Proteome Project. Nat Struct Biol 6:11-2
|
CABM
Home
Top |
