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