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A novel NMR method for determining the interfaces of large protein–protein complexes

Nature Structural Biology volume 7, pages 220–223 (2000)Cite this article

Abstract

Identification of the interfaces of large (Mr > 50,000) protein–protein complexes in solution by high resolution NMR has typically been achieved using experiments involving chemical shift perturbation and/or hydrogen-deuterium exchange of the main chain amide groups of the proteins. Interfaces identified using these techniques, however, are not always identical to those revealed using X-ray crystallography. In order to identify the contact residues in a large protein–protein complex more accurately, we developed a novel NMR method that uses cross-saturation phenomena in combination with TROSY detection in an optimally deuterium labeled system.

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Figure 1: Principles of the cross-saturation method.
Figure 2: Pulse scheme for the cross-saturation method.
Figure 3: The results of the cross-saturation experiment.
Figure 4: Plots of the intensity ratios of the crosspeaks in the cross-saturation experiments.
Figure 5: Comparison of the binding sites of the FB–Fc complex.

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Acknowledgements

We thank F. Delaglio for providing scripts for spectra analysis and M. Nakasako for useful discussions.

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Authors and Affiliations

  1. Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, 113-0033, Tokyo, Japan

    Hideo Takahashi, Tamiji Nakanishi, Keiichiro Kami & Ichio Shimada

  2. Water Research Institute, Sengen, Tsukuba, 305-0047, Ibaraki, Japan

    Yoji Arata

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  1. Hideo Takahashi
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Correspondence to Ichio Shimada.

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Takahashi, H., Nakanishi, T., Kami, K. et al. A novel NMR method for determining the interfaces of large protein–protein complexes. Nat Struct Mol Biol 7, 220–223 (2000). https://doi.org/10.1038/73331

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