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Protein fold determined by paramagnetic magic-angle spinning solid-state NMR spectroscopy

Nature Chemistry volume 4, pages 410–417 (2012)Cite this article

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Abstract

Biomacromolecules that are challenging for the usual structural techniques can be studied with atomic resolution by solid-state NMR spectroscopy. However, the paucity of distance restraints >5 Å, traditionally derived from measurements of magnetic dipole–dipole couplings between protein nuclei, is a major bottleneck that hampers such structure elucidation efforts. Here, we describe a general approach that enables the rapid determination of global protein fold in the solid phase via measurements of nuclear paramagnetic relaxation enhancements (PREs) in several analogues of the protein of interest containing covalently attached paramagnetic tags, without the use of conventional internuclear distance restraints. The method is demonstrated using six cysteine–EDTA–Cu2+ mutants of the 56-residue B1 immunoglobulin-binding domain of protein G, for which ~230 longitudinal backbone 15N PREs corresponding to distances of ~10–20 Å were obtained. The mean protein fold determined in this manner agrees with the X-ray structure with a backbone atom root-mean-square deviation of 1.8 Å.

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Figure 1: Determination of longitudinal backbone amide 15N PREs in Cys–EDTA–Cu2+ GB1 mutants by solid-state NMR spectroscopy.
Figure 2: Results of idealized preliminary structure calculations for GB1 in the absence and presence of solid-state NMR longitudinal 15N PRE restraints.
Figure 3: De novo calculation of the GB1 fold using solid-state NMR longitudinal 15N PRE restraints.

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Acknowledgements

This research was supported by the National Science Foundation (CAREER award MCB-0745754 to C.P.J.). C.D.S. was supported by the National Institutes of Health Intramural Research Program of the Center for Information Technology. The GB1 plasmid was kindly provided by A.M. Gronenborn.

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Author notes

  1. Jonathan J. Helmus

    Present address: Present address: Department of Molecular, Microbial, and Structural Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, Connecticut 06030, USA,

  2. Ishita Sengupta and Philippe S. Nadaud: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Chemistry, The Ohio State University, 100 West 18th Avenue, Columbus, 43210, Ohio, USA

    Ishita Sengupta, Philippe S. Nadaud, Jonathan J. Helmus & Christopher P. Jaroniec

  2. Division of Computational Bioscience, Center for Information Technology, Building 12A, National Institutes of Health, Bethesda, 20892, Maryland, USA

    Charles D. Schwieters

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Contributions

C.P.J. designed the research. I.S. and P.S.N. prepared the samples. I.S., P.S.N. and J.J.H. recorded and analysed the NMR data. C.D.S. and C.P.J. performed the structure calculations. C.D.S. and C.P.J. wrote the paper.

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Correspondence to Christopher P. Jaroniec.

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Sengupta, I., Nadaud, P., Helmus, J. et al. Protein fold determined by paramagnetic magic-angle spinning solid-state NMR spectroscopy. Nature Chem 4, 410–417 (2012). https://doi.org/10.1038/nchem.1299

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