Nuclear Magnetic Resonance (NMR) Spectroscopy of Proteins

Abstract

Nuclear magnetic resonance (NMR) spectroscopy enables the determination of three‐dimensional protein structures at atomic resolution under near‐physiological conditions in solution. In structural biology, NMR complements X‐ray crystallography, which provides similar information on proteins in single crystals.

Keywords: nuclear magnetic resonance; protein structure; protein dynamics; resonance assignments

Figure 1.

Diagram outlining the course of a protein structure determination by NMR in solution (see text).

Figure 2.

Three‐dimensional 15N‐correlated [1H,1H]‐NOESY spectrum of uniformly 15N‐labelled E. coli glutaredoxin.

Figure 3.

(a) Sequential resonance assignment based on sequential 1H–1H NOEs. In the dipeptide segment –Ala–Val– the dotted lines indicate 1H–1H relations that can be established by scalar, through‐bond spin–spin couplings seen in homonuclear 1H NMR experiments. The broken arrows indicate pairs of protons in sequentially neighbouring residues that can be related by 1H–1H NOEs that manifest short sequential distances dαN (between αCH and the amide proton of the following residue) and dNN (between the amide protons of neighbouring residues). (b) Segment of a uniformly 13C/15N‐labelled polypeptide chain with indication of the scalar spin–spin couplings (in Hz) that provide the basis for obtaining sequential assignments by triple‐resonance experiments (see text).

Figure 4.

Protein structure determination based on upper distance constraints measured by 1H–1H nuclear Overhauser effects (NOEs). Part of a 2D [1H,1H]‐NOESY spectrum of the Antennapedia homeodomain is shown, and at the top there is a line, N→C that represents a polypeptide chain. Two 1H atoms of the polypeptide are identified by circles, and straight arrows point from the corresponding peaks on the diagonal in the NOESY spectrum to these protons (these arrows represent the result of the sequential assignment). The presence of a NOESY cross‐peak outside of the diagonal, which is connected with the two aforementioned diagonal positions by a vertical and a horizontal line, shows that these two protons are separated by a short distance (≤ 4.0 Å), and therefore the observation of this single NOE imposes a looped structure on the polypeptide chain (top right). Each additional NOESY cross‐peak defines a similar loop. The NMR structure corresponds to the molecular geometry that contains simultaneously all the loops defined by the NOESY data.

Figure 5.

Stereo view of the polypeptide backbone of 20 energy‐refined conformers selected to represent the NMR solution structure of the 60‐residue tick anticoagulant protein from Ornithodoros moubata. The numbers identify selected sequence positions.

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References

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Further Reading

Cavanagh J, Fairbrother WJ, Palmer AG III and Skelton NJ (1996) Protein NMR Spectroscopy, Principles and Practice. New York: Academic Press.

Ernst RR, Bodenhausen G and Wokaun A (1987) Principles of Nuclear Magnetic Resonance in One and Two Dimensions. Oxford: Clarendon.

Jardetzky O and Roberts GCK (1981) NMR in Molecular Biology. New York: Academic Press.

Kay LE and Gardner KH (1997) Solution NMR spectroscopy beyond 25 kDa. Current Opinion in Structural Biology 7: 722–731.

Otting G, Liepinsh E and Wüthrich K (1991) Protein hydration in aqueous solution. Science 254: 974–980.

Wider G and Wüthrich K (1999) NMR with large molecules and multimolecular assemblies in solution. Current Opinion in Structural Biology 9: 594–601.

Wishart DS, Sykes BD and Richards FM (1991) Relationship between nuclear‐magnetic‐resonance chemical shift and protein secondary structure. Journal of Molecular Biology 222: 311–333.

Wüthrich K (1970) Structural studies of hemes and hemoproteins by nuclear magnetic resonance spectroscopy. Structure and Bonding 8: 53–121.

Wüthrich K (1995) NMR – this other method for protein and nucleic acid structure determination. Acta Crystallographica D 51: 249–270.

Wüthrich K (1995) NMR in Structural Biology. Singapore: World Scientific.

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How to Cite close
Wüthrich, Kurt(Apr 2001) Nuclear Magnetic Resonance (NMR) Spectroscopy of Proteins. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0003103]