Structure determination of protein complexes by NMR.
This chapter describes nuclear magnetic resonance (NMR) methods that can be used to determine the structures of protein complexes. Many of these techniques are also applicable to other systems (e.g., protein-nucleic acid complexes). In the first section, we discuss methodologies for optimizing the sample conditions for the study of complexes. This is followed by a description of the methods that can be used to map interfaces when a full structure determination of the complex is not appropriate or not possible. We then describe experimental approaches for resonance assignment in complexes, these are essentially the same as those for isolated proteins. Subheading 6. describes the different types of so-called X-filtered NMR experiments that have been devised to separate and selectively observe either inter- or intramolecular structural information. These filtered NMR experiments are then exploited in the experimental strategies for structure determination of either protein complexes or homodimeric proteins. This is followed by a description of the calculation of their structures. Finally, we present case studies from three projects carried out in our laboratory, where we successfully used the methods presented in this chapter.
Nietlispach
Daniel
D
Department of Biochemistry, University of Cambridge, UK.
Mott
Helen R
HR
Stott
Katherine M
KM
Nielsen
Peter R
PR
Thiru
Abarna
A
Laue
Ernest D
ED
D000818:Animals; D019204:GTP-Binding Proteins / Q000737:chemistry; D006657:Histones / Q000737:chemistry; D046911:Macromolecular Substances; D051379:Mice; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D054462:p21-Activated Kinases
D006657:Histones; D046911:Macromolecular Substances; D009841:Oligonucleotides; D010455:Peptides; D011506:Proteins; D017346:Protein Serine-Threonine Kinases; D054462:p21-Activated Kinases; D019204:GTP-Binding Proteins