Место издания:Kazan Federal Universsity Kazan, Russian Federation
Первая страница:23
Последняя страница:23
Аннотация:Studies of the molecular recognition processes are essential for understanding of the mechanism of numerous biochemical processes in a live organism, as well as for developing new rational approaches of the design of new biologically active compounds. Developments of new hardware and techniques in the last decades have strengthened position of NMR spectroscopy as one of the most powerful and informative biophysical methods of biological macromolecule structure determination and elucidation of protein-ligand interactions. NMR spectroscopy gives ample opportunities for studying protein-ligand interactions. Some of such opportunities will be illustrated in the present report with the results of our ongoing studies. The report will briefly describe some of the basic principles involved in studies of protein–ligand interactions and identify the range of possible applications.
One concept assumes structure determination of a protein-ligand complex in solution using the methods of NMR spectroscopy. Moreover NMR techniques allow also to identify specific protein-ligand interactions and to study dynamic properties of a complex in broad timescale of internal motions. This approach will be illustrated by the results of studies of highly specific interaction of bacterial dihydrofolate reductase with its inhibitor - an antibacterial drug trimethoprim, and also by the results of our ongoing studies of zinc ion interaction with the metal binding domain of β-amyloid peptide, involved in the development of the Alzheimer’s disease.
Another “chemical shift mapping” approach is based on the analysis of protein chemical shift changes caused by interaction with ligands. Interacting protein residues can be identified and mapped on the surface of a known protein structure. This will be illustrated by the studies of mRNA stop codon recognition by the human translation termination factor eRF1. Similar by the concept approach was also used for the determination of the interaction interface of the middle domain of human eRF1 with the large subunit of eukaryotic ribosome.
Third approach assumes use of NMR spectroscopy as a tool of qualitative screening of pharmaceutical libraries to assess specific binding of the ligands to target protein. In contrast to the approaches mentioned above, NMR screening techniques are based on the detection of spectral parameters of ligands instead of protein. Therefore it increases sensitivity of such NMR techniques, allows reducing concentration of studied protein and removes need of its highly expensive isotope labeling. Opportunities of modern NMR screening techniques will be illustrated with the results of our ongoing studies aimed at the design of the inhibitors of methionine γ-lyase – an important potential antibacterial drug target.