Biological NMR Centre, University of Leicester, Leicester LE1 9HN, UK
NMR spectroscopy can provide information on many different aspects of protein-ligand interactions, ranging from the determination of the complete structure of the complex to focusing on selected features of the interactions by using "reporter groups" on the ligand or the protein (Feeney & Birdsall, 1993; Lian et al., 1994). In this talk I will discuss recent work on several enzymes involved in drug action or drug metabolism to illustrate the kind of information which can be obtained about the active sites of enzymes and about substrate binding:
[ Dihydrofolate reductase (18kD): studies of mutants, and of the effects of coenzyme binding on protein dynamics (Basran et al., 1995; Casarotto et al., 1995).
[ TEM [[beta]]-lactamase (30kD): determination of pK values of active site residues in the study of catalytic mechanisms (Damblon et al., 1995).
[ Cytochrome P450 (51kD): use of paramagnetic relaxation to "dock" substrates into the binding site (Modi et al., 1995).
[ Chloramphenicol acetyltransferase (75kD): use of transferred NOEs to determine the conformation of a bound substrate (Barsukov et al., 1995).
Barsukov, I.L., Lian, L.-Y., Ellis, J., Shaw, W.V., and Roberts, G.C.K. (1995) submitted.
Basran, J., Casarotto, M.A., Barsukov, I.L., and Roberts, G.C.K. (1995) Biochemistry 34, 2872-2882.
Casarotto, M.A., Sze, K.-H., Barsukov, I.L., and Roberts, G.C.K. (1995) submitted.
Damblon, C., Raquet, X., Lian, L.-Y., Lamotte-Brasseur, J., Fonzé, E., Charlier, P., Roberts, G.C.K., and Frère, J.-M. (1995) submitted.
Feeney, J., and Birdsall, B. (1993) In: NMR of Biological Macromolecules, Ed. Roberts, G.C.K. IRL Press at Oxford University Press.
Lian, L.-Y., Barsukov, I.L., Sutcliffe, M.J., Sze, K.H., and Roberts, G.C.K. (1994) Meth. Enzymol. 239, 657-700.
Modi, S., Primrose, W.U., Boyle, J.M.B., Gibson, C.F., Lian, L.-Y., and Roberts, G.C.K. (1995) Biochemistry, in press.