Genetic Methods in High-Resolution NMR Studies of Proteins
Many biomolecules have long rotational correlation times and thus give rise to broad NMR resonances in solution. A variety of strategies have been devised to overcome this limitation. Resolution enhancement algorithms have permitted the study of flexible regions of structures as large as Tobacco Mosaic Virus (1–3); one-dimensional and two-dimensional pulse sequences may also be used to selectively observe flexible spin systems (4,5). In addition, the introduction of nuclear spin labels has facilitated the identification of individual residues (6–9). In this paper we describe how the techniques of molecular biology may be employed to facilitate the NMR study of large proteins. Illustrative examples will be taken from our studies of the bacteriophage λ repressor (10–12).
KeywordsTobacco Mosaic Virus Broad Resonance Major Groove Methyl Resonance Aliphatic Region
- 10.M. A. Weiss, M. Karplus, D. J. Patel, and R. T. Sauer, Solution NMR Studies of Intact λ Repressor, J. Biomol. Struc. Dyn. 1:150 (1983).Google Scholar
- 12.M. A. Weiss, M. Karplus, and R. T. Sauer, Complete Assignment of the Aromatic Resonances of λ Repressor, manuscript in preparation.Google Scholar