protein by NMR seem feasible (Gordon & Wu$thrich, 1978; Wagner & Wu$thrich, 1979). .... assignments are made during a later stage of the assignment process. .... for labeling RNA have been developed and it is now possible to work with larger and more ..... already assigned, the resulting NOE list is not as reliable.
Quarterly Reviews of Biophysics 33, 1 (2000), pp. 29–65 # 2000 Cambridge University Press
Printed in the United Kingdom
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NMR spectroscopy : a multifaceted approach to macromolecular structure Ann E. Ferentz and Gerhard Wagner Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Cambridge, MA USA
1. Introduction
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2. Landmarks in NMR of macromolecules 32 2.1 Protein structures and methods development 32 2.1.1 Sequential assignment method 32 2.1.2 Triple-resonance experiments 34 2.1.3 Structures of large proteins 36 2.2 Protein–nucleic acid complexes 37 2.3 RNA structures 38 2.4 Membrane-bound systems 39 3. NMR spectroscopy today 40 3.1 State-of-the-art structure determination 41 3.2 New methods 44 3.2.1 Residual dipolar couplings 44 3.2.2 Direct detection of hydrogen bonds 44 3.2.3 Spin labeling 45 3.2.4 Segmental labeling 46 3.3 Protein complexes 47 3.4 Mobility studies 50 3.5 Determination of time-dependent structures 52 3.6 Drug discovery 53 4. The future of NMR 54 4.1 The ease of structure determination 54 4.2 The ease of making recombinant protein 55 4.3 Post-translationally modified proteins 55 4.4 Approaches to large and/or membrane-bound proteins 56 4.5 NMR in structural genomics 56 4.6 Synergy of NMR and crystallography in protein structure determination 56 5. Conclusion
57
6. Acknowledgements
57
7. References 57
1. Introduction Since the publication of the first complete solution structure of a protein in 1985 (Williamson et al. 1985), tremendous technological advances have brought nuclear magnetic resonance
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Ann E. Ferentz and Gerhard Wagner
Table 1. Numbers of structures in the Protein Data Bank as of January 2000 Type of macromolecule
NMR
X-ray diffraction & other
Proteins, peptides, viruses Protein–nucleic acid complexes Nucleic acids Carbohydrates Total
1471 62 286 4 1823
8488 411 473 14 9386
Adapted from the RCSB Protein Data Bank web site.
500 (a) NMR depositions in the PDB 450
Number of structures
400 350 300 250 200 150 100 50 0
1986
1987
1988
1989
1990
1991
1992 Year
1993
1991 1992 Year
1993
1994
1995
1996
1997
1998
2000 (b) Crystallographic depositions in the PDB 1800
Number of structures
1600 1400 1200 1000 800 600 400 200 0
1986
1987
1988 1989
1990
1994
1995
1996 1997 1998
Fig. 1. Numbers of entries in the Protein Data Bank as of December 1999. (a) Number of depositions of NMR structures each year. (b) Number of X-ray crystal structures deposited each year.
NMR spectroscopy : a multifaceted approach to macromolecular structure
