title: Two-point anchoring of a lanthanide-binding peptide to a target protein enhances the paramagnetic anisotropic effect.
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.
authors.affiliation: Graduate School of Life Science, Hokkaido University, Sapporo, 001-0021, Japan.
authors.forename: Tomohide
authors.initials: T
authors.lastname: Saio
authors.forename: Kenji
authors.initials: K
authors.lastname: Ogura
authors.forename: Masashi
authors.initials: M
authors.lastname: Yokochi
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Kobashigawa
authors.forename: Fuyuhiko
authors.initials: F
authors.lastname: Inagaki
mesh_terms: D028581:Lanthanoid Series Elements / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*
chemical_list: D028581:Lanthanoid Series Elements; D011506:Proteins