Two-point anchoring of a lanthanide-binding peptide to a target protein enhances the paramagnetic anisotropic effect
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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.
KeywordsLanthanide binding peptide tag Two-point anchoring Paramagnetic NMR Pseudo-contact shift Residual dipolar coupling
- Banci L, Bertini I, Huber JG, Luchinat C, Rosato A (1998) Partial orientation of oxidized and reduced cytochrome b5 at high magnetic fields: magnetic susceptibility anisotropy contributions and consequences for protein solution structure determination. J Am Chem Soc 120:12903–12909CrossRefGoogle Scholar
- DeLano WL (2002) The PyMOL molecular graphics system. Palo Alto, CAGoogle Scholar