title: NMR analysis of viral protein structures.
abstract: Nuclear magnetic resonance (NMR) spectroscopy is a powerful tool to study the three-dimensional structure of proteins and nucleic acids at atomic resolution. Since the NMR data can be recorded in solution, conditions such as pH, salt concentration, and temperature can be adjusted so as to closely mimic the biomacromolecules natural milieu. In addition to structure determination, NMR applications can investigate time-dependent phenomena, such as dynamic features of the biomacromolecules, reaction kinetics, molecular recognition, or protein folding. The advent of higher magnetic field strengths, new technical developments, and the use of either uniform or selective isotopic labeling techniques, currently allows NMR users the opportunity to investigate the tertiary structure of biomacromolecules of approximately 50 kDa. This chapter will outline the basic protocol for structure determination of proteins by NMR spectroscopy. In general, there are four main stages: (i) preparation of a homogeneous protein sample, (ii) the recording of the NMR data sets, (iii) assignment of the spectra to each NMR observable atom in the protein, and (iv) generation of structures using computer software and the correctly assigned NMR data.
authors: Department of Chemistry and School of Biological Sciences, The University of Auckland, Science Centre, 23 Symonds Street, Auckland, New Zealand.
authors: Andrew J
authors: AJ
authors: Dingley
authors: Inken
authors: I
authors: Lorenzen
authors: Joachim
authors: J
authors: Grötzinger
mesh_terms: D003001:Cloning, Molecular; D018076:DNA, Complementary; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008958:Models, Molecular; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D012996:Solutions; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D018076:DNA, Complementary; D007202:Indicators and Reagents; D011994:Recombinant Proteins; D012996:Solutions; D014764:Viral Proteins