Gaetano Montelione

New NMR methods development, molecular recognition, growth factors, protein: nucleic acid complexes,
DNA and RNA structure, molecular dynamics, protein design.

The general aim of our research is to use NMR spectroscopy as a tool for protein engineering and rational drug design. We develop new methods for protein solution structure determination and apply these techniques to proteins of pharmaceutical or medical interest. The combined methods of site-directed mutagenesis, NMR spectroscopy, and conformational energy calculations are being used to (1) determine three dimensional structures of small proteins in solution, (2) determine the structures of protein-protein, protein-receptor, and protein-nucleic acid complexes, (3) characterize effects of amino acid substitutions on protein structure, stability, and provide information for rational drug design, and (5) study the molecular mechanism by which proteins fold into their biologically-active conformations. We are currently working on structure determination and refinement of several protein growth factors including epidermal and type-a transforming growth factors, insulin-like growth factors, and heparin-binding growth factor. Several DNA-and-RNA-binding proteins are also being studied by NMR. We have recently determine solution structures of an IgG-binding domain of staphylococcal Protein A and of an RNA-binding protein from E. coli which is overproduced in response to cold shock (Cold Shock Protein A). We have characterized structural changes in these molecules which are required for binding to IgG proteins or to nucleic acids. Nuclear relaxation time measurements are used to characterize intramolecular motion in these small proteins. This research has important implications in the field of protein physical chemistry, molecular design, receptor-ligand interactions, and oncogenesis.

Recent Publications Montelione, G.T., Lyons, B.A., Emerson, S.D., Tashiro, M. (1993). An efficient triple resonance experiment using carbon-13 isotropic mixing for determining sequence-specific resonance assignments of isotopically enriched proteins, J. Amer Chem. Soc. 114: 10974-10975. Moy, F.J., Li, Y.-C., Winkler, M.E., Scheraga, H.A., Montelelione, G.T. (1993). Solution structure of human type-a transforming growth factor determine by heteronuclear NMR spectroscopy and refined by energy minimization, Biochemistry 32: 7334-7353. Lyons, B.A., Tashiro, M., Cedergren, L., Nilsson, B., Montelione, G.T. (1993). An improved strategy for determining sequence-specific resonance assignments of isotopically enriched proteins and its applications on an engineering domain of staphylococcal Protein A, Biochemistry 32: 7839-7845. Newkirk, K., Feng, W., Jiang, W., Tejero, R., Emerson, S.D., Inouye, M., Montelione, G.T. (1994). Solution NMR structure of the major cold-shock protein (CspA) from Escherichia coli: Identification of a binding epitope for single-stranded DNA, Proc. Natl. Acad. Sci. U.S.A. 91: 5114-5118.