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Two-point anchoring of a lanthanide-binding peptide to a target protein enhances the paramagnetic anisotropic effect

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Abstract

Paramagnetic lanthanide ions fixed in a protein frame induce several paramagnetic effects such as pseudo-contact shifts and residual dipolar couplings. These effects provide long-range distance and angular information for proteins and, therefore, are valuable in protein structural analysis. However, until recently this approach had been restricted to metal-binding proteins, but now it has become applicable to non-metalloproteins through the use of a lanthanide-binding tag. Here we report a lanthanide-binding peptide tag anchored via two points to the target proteins. Compared to conventional single-point attached tags, the two-point linked tag provides two to threefold stronger anisotropic effects. Though there is slight residual mobility of the lanthanide-binding tag, the present tag provides a higher anisotropic paramagnetic effect.

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Authors and Affiliations

  1. Graduate School of Life Science, Hokkaido University, Sapporo, 001-0021, Japan

    Tomohide Saio

  2. Laboratory of Structural Biology, Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo, 001-0021, Japan

    Tomohide Saio, Kenji Ogura, Masashi Yokochi, Yoshihiro Kobashigawa & Fuyuhiko Inagaki

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  1. Tomohide Saio
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  2. Kenji Ogura
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  3. Masashi Yokochi
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  4. Yoshihiro Kobashigawa
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  5. Fuyuhiko Inagaki
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Correspondence to Fuyuhiko Inagaki.

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Saio, T., Ogura, K., Yokochi, M. et al. Two-point anchoring of a lanthanide-binding peptide to a target protein enhances the paramagnetic anisotropic effect. J Biomol NMR 44, 157–166 (2009). https://doi.org/10.1007/s10858-009-9325-z

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  • DOI: https://doi.org/10.1007/s10858-009-9325-z

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