NMR Studies of Protein-Ligand Interactions: Dihydrofolate Reductase
The study of protein-ligand interactions has proved to be one of the more fruitful applications of high-resolution nmr to biological problems. The specific recognition of small molecules by proteins is fundamental to a wide range of biological phenomena, from enzyme action and its regulation, through the control of gene expression to the action of hormones and neurotransmitters. Nmr can provide a variety of information on both structural and dynamic aspects of the recognition process in the solution state. In this article I shall describe some of the different kinds of Information which can be obtained, drawing illustrations from work on the enzyme dlhydrofolate reductase, which we have been studying for a number of years. A broader view of the wide range of experiments which have been done on other systems can be found in ref. 1, while fuller references to the work on dlhydrofolate reductase are given in ref. 2. Operationally, two kinds of experiments can be distinguished, depending on whether one studies resonances of the small molecule or of the protein, and examples of both will be discussed. First, however, we must consider, briefly, the effects of chemical exchange processes on nmr spectra and their interpretation.
KeywordsChemical Shift Free State Ligand Concentration Dihydrofolate Reductase Dissociation Rate Constant
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