Protein alignment by a coexpressed lanthanide-binding tag for the measurement of residual dipolar couplings.
Title | Protein alignment by a coexpressed lanthanide-binding tag for the measurement of residual dipolar couplings. |
Publication Type | Journal Article |
Year of Publication | 2003 |
Authors | Wöhnert J, Franz KJ, Nitz M, Imperiali B, Schwalbe H |
Journal | J Am Chem Soc |
Volume | 125 |
Pagination | 13338–13339 |
Date Published | Nov |
Accession Number | 72 |
Abstract | A protein fusion construct of human ubiquitin with an N-terminal lanthanide binding tag (LBT) enables observation of long-range orientational restraints in solution NMR from residual dipolar couplings (RDCs) due to paramagnetic alignment of the protein. The paramagnetic lanthanide ions Tb3+, Dy3+, and Tm3+ are shown to bind to the LBT and induce different alignment tensors, in agreement with theory. RDCs, measured relative to the diamagnetic Lu3+, range from -7.6 to 5.5 Hz for Tb3+ and -6.6 to 6.1 Hz for Dy3+, while an opposite alignment tensor is observed for Tm3+ (4.5 to -2.9 Hz) at 800 MHz. Experimental RDCs are in excellent agreement with those predicted on the basis of the X-ray structure of the protein. |
URL | http://dx.doi.org/10.1021/ja036022d |
DOI | 10.1021/ja036022d |