Protein alignment by a coexpressed lanthanide-binding tag for the measurement of residual dipolar couplings.

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Wöhnert, J; Franz, KJ; Nitz, M; Imperiali, B; Schwalbe, H

Published Date

  • November 2003

Published In

Volume / Issue

  • 125 / 44

Start / End Page

  • 13338 - 13339

PubMed ID

  • 14583012

Electronic International Standard Serial Number (EISSN)

  • 1520-5126

International Standard Serial Number (ISSN)

  • 0002-7863

Digital Object Identifier (DOI)

  • 10.1021/ja036022d


  • eng