Generation of pseudocontact shifts in proteins with lanthanides using small "clickable" nitrilotriacetic acid and iminodiacetic acid tags

Choy-Theng Loh, Bim Graham, Elwy H Abdelkader, Kellie L Tuck, Gottfried Otting

Research output: Contribution to journalArticleResearchpeer-review

19 Citations (Scopus)

Abstract

Pseudocontact shifts (PCS) induced by paramagnetic lanthanide ions provide unique long-range structural information in nuclear magnetic resonance (NMR) spectra, but the site-specific attachment of lanthanide tags to proteins remains a challenge. Here we incorporated p-azido-phenylalanine (AzF) site-specifically into the proteins ubiquitin and GB1, and ligated the AzF residue with alkyne derivatives of small nitrilotriacetic acid and iminodiacetic acid tags using the CuI-catalysed "click" reaction. These tags form lanthanide complexes with no or only a small net charge and produced sizeable PCSs with paramagnetic lanthanide ions in all mutants tested. The PCSs were readily fitted by single magnetic susceptibility anisotropy tensors. Protein precipitation during the click reaction was greatly alleviated by the presence of 150 mM NaCl.
Original languageEnglish
Pages (from-to)5084-5092
Number of pages9
JournalChemistry - A European Journal
Volume21
Issue number13
DOIs
Publication statusPublished - 2015

Keywords

  • click chemistry
  • lanthanide-binding tags
  • NMR spectroscopy
  • p-azido-L-phenylalanine
  • pseudocontact shifts

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