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A chiral lanthanide tag for stable and rigid attachment to single cysteine residues in proteins for NMR, EPR and time-resolved luminescence studies
journal contribution
posted on 2021-08-10, 08:57 authored by Iresha D Herath, Colum Breen, Sarah Hewitt, Thomas Berki, Ahmad Kassir, Charlotte Dodson, Martyna Judd, Shereen Jabar, Nicholas Cox, Gottfried Otting, Stephen ButlerStephen ButlerA lanthanide-binding tag site-specifically attached to a protein presents a tool to probe the protein by multiple spectroscopic techniques, including nuclear magnetic resonance, electron paramagnetic resonance and time-resolved luminescence spectroscopy. Here a new stable chiral LnIII tag, referred to as C12, is presented for spontaneous and quantitative reaction with a cysteine residue to generate a stable thioether bond. The synthetic protocol of the tag is relatively straightforward, and the tag is stable for storage and shipping. It displays greatly enhanced reactivity towards selenocysteine, opening a route towards selective tagging of selenocysteine in proteins containing cysteine residues. Loaded with TbIII or TmIII ions, the C12 tag readily generates pseudocontact shifts (PCS) in protein NMR spectra. It produces a relatively rigid tether between lanthanide and protein, which is beneficial for interpretation of the PCSs by single magnetic susceptibility anisotropy tensors, and it is suitable for measuring distance distributions in double electron-electron resonance experiments. Upon reaction with cysteine or other thiol compounds, the TbIII complex exhibits a 100-fold enhancement in luminescence quantum yield, affording a highly sensitive turn-on luminescence probe for time-resolved FRET assays and enzyme reaction monitoring.
Funding
Luminescent Host Molecules for Multisite Recognition of Polyphosphate Anions EP/S032339/1
Wellcome Trust (204500/Z/16/Z)
Laureate Fellowship, Australian Research Council. Grant Number: FL170100019
Australian Research Council. Grant Number: DP170100162
Centre of Excellence for Innovations in Peptide & Protein Science, Australian Research Council. Grant Number: CE200100012
History
School
- Science
Department
- Chemistry
Published in
Chemistry: A European JournalVolume
27Issue
51Pages
13009-13023Publisher
WileyVersion
- VoR (Version of Record)
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© The authorsPublisher statement
This is an Open Access Article. It is published by Wiley under the Creative Commons Attribution 4.0 Unported Licence (CC BY). Full details of this licence are available at: http://creativecommons.org/licenses/by/4.0/Acceptance date
2021-06-09Publication date
2021-07-29Copyright date
2021ISSN
0947-6539eISSN
1521-3765Publisher version
Language
- en
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Deposit date: 10 August 2021Usage metrics
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