Structure Determination of Protein Complexes by NMR

Structure Determination of Protein Complexes by NMR
D. Nietlispach, H.R. Mott, K.M. Stott, P.R. Nielsen, A. Thiru & E.D. Laue
The Department of Biochemistry, University of Cambridge

Introduction
As the structures of more proteins and domains are solved by structural genomics projects, the future of structural biology will be oriented more toward the study of macromolecular complexes. Since so many biological processes are mediated by interactions between proteins, it is important to study them at a molecular level. The study of protein-protein interactions also has applications in a number of therapeutic areas, particularly drug design, but also in understanding the molecular basis of human diseases. NMR spectroscopy is highly suited to investigate molecular interactions between proteins or proteins and peptides at close to physiological conditions. Such interactions may be of variable strength and sample a variety of timescales. While it is possible to obtain a high resolution structure of a complex of up to 40 kDa, it is also feasible to map the interface of much larger complexes. The majority of structural studies of molecular assemblies have been cases where the dissociation constant of the complex is sub-micromolar; a situation referred to as tight or strong binding. Under such conditions, a full structure determination of the complex is feasible. For weaker binding cases, structural information can be obtained for a ligand in the bound state and the ligand is then modelled onto the surface of the known protein structure.

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